Next Generation Sequencing Catalog Scientific NGS Catalog 2014... · 2015-07-02 · Introduction 2 Bioo Scientific Bioo Scientific is a global life science company headquartered in

Next Generation Sequencing Catalog2014-2015

®

New RapidDNA-Seq Kitsee page 06

New 16S V4Amplicon-Seq Kit

see page 22

NewqRNA-Seq Kitsee page 40

Intro

duct

ion

2

Bioo ScientificBioo Scientific is a global life science company headquartered in Austin, TX, committed to providing innovative products and superior cus-tomer service. We redefine the limits of what’s possible in Next Generation Sequencing, affording unprecedented throughput and efficiency for faster, higher-quality results. We are continually adding to our product line to increase the sensitivity, flexibility and speed of NGS experi-ments, driving breakthroughs in the field of genomics.

Quality and delivery time are important considerations when deciding upon a vendor for your library preparations kits and reagents. Our focus has been developing enzymes (ligases and polymerases) that deliver the highest efficiency to achieve robust results while minimizing library prep associated bias. We’ve designed our flagship library preparation solutions to optimize adapter ligation efficiency, generating a greater number of unique reads, higher sequence diversity, and greater coverage. Bioo Scientific also has an optimized inventory management sys-tem to ensure overnight delivery.

In recognition of our achievements, Bioo Scientific has also been honored to receive the following awards:

2013 – Greater Austin Business Technology Award2013 – APEX Award for Publication Excellence – Print Ads & Advertorials2011 – Greater Austin Business Export Award2009 – Tech Innovator of the Year Award – Biotech/Medical Division

Table of Contents

3

DNA-Seq Library Prep - Illumina® Compatible

DNA-Seq Library Prep - Ion PGM™ & Proton™ Compatible

DNA-Seq Library Prep - SOLiD™ Compatible

RNA-Seq Library Prep

Next-Gen Sequencing Automation

DNA Fragmentation & Nucleic Acid Isolation

Ordering Instructions

04

28

32

38

52

56

62

Table of Contents

4 BiooNGS.com The NGS Experts™®

5BiooNGS.com The NGS Experts™®

DNA-Seq Library PrepIllumina® Compatible

BiooNGS.com The NGS Experts™®6

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NEXTflex™ Rapid DNA Sequencing Kitsfor Illumina®

Bioo Scientific’s NEXTflex™ Rapid DNA-Seq Kit allows researchers to build sequencing-ready libraries for sequencing on Illumina® sequencing platforms, from as little as 1 ng of DNA, in two hours or less. This kit is ideal for use with genomic DNA, FFPE samples, ChIP DNA and low input clinical samples.

The NEXTflex Rapid DNA-Seq Kit is highly flexible in terms of sample requirements, accommodating a 3-log range of input DNA amounts from 1 ng to 1 µg. A bead-based, gel-free size selection protocol eliminates the need for agarose gels in size selection, and the availability of up to 96 unique adapter barcodes facilitates high-throughput applications. The NEXTflex Rapid DNA-Seq Kit incorporates Enhanced Adapter Ligation Technology, which facilitates ligation of long adapters, resulting in longer and more diverse sequencing reads. Bioo Scientific’s NEXTflex Ligation and Polymerase reaction mixes ensure the highest quality libraries for superior performance.

For multiplexing, The NEXTflex™ Adapters are long, annealed adapters containing indexed sequences that offer an improved multiplexing work-flow and flexible setup. These barcodes can be used with single, paired-end and multiplex reads. The NEXTflex Rapid DNA-Seq Kit is designed to be used with the NEXTflex™ and NEXTflex-96™ DNA Barcodes for 10 ng or more of starting material or the NEXTflex™ and NEXTflex-96™ ChIP-Seq Barcodes for less than 10 ng of starting material. These barcodes are available in sets of 6, 12, 24, 48 and 96 unique indexed adapters.

• Flexible amounts of input DNA from 1 ng to 1 µg• Fast workflow requiring 2 hours or less, with minimal hands-on time• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads• Creates libraries with less bias than alternative methods• Automation-friendly workflow is compatible with liquid handlers• Flexible barcode options – Up to 96 unique barcodes• Compatible with Illumina® HiSeq, MiSeq and GAIIx platforms• Prepare single, paired-end and multiplexed genomic DNA libraries• Bulk pricing available to meet research goals

End Polishing

Adapter Ligation

PCR

1 ng – 1 µg

2 hours or less

For automated solutions see page 54.

Figure 1. The NEXTflex Rapid DNA-Seq Kit features a stream-lined library prep protocol to help you meet your research goals faster than you thought possible.

Con

cent

ratio

n (p

M)

Library Samples

16.00

14.00

12.00

10.00

8.00

6.00

4.00

2.00

0.00

NEXTflex, 6 cycles

Phusion, 6 cycles

NEXTlfex, 8 cycles

Phusion, 8 cycles

Library Samples (nM) STDEV

NEXTflex, 6 cycles 5.65 2.03

Phusion, 6 cycles 3.18 1.86

NEXTflex, 8 cycles 12.72 1.11

Phusion, 8 cycles 6.31 3.69

Figure 2. Higher yield obtained by NEXTflex Rapid DNA-Seq Kit com-pared to competitor kit. The final nanomolar concentration measurements of each triplicate PCR condition by qPCR. Each triplicate Ct measurement was averaged, converted to pM (then nM) using the standard curve, determined, then plotted with standard deviation against the other PCR conditions.

BiooNGS.com The NGS Experts™® 7

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100.0

90.0

80.0

70.0

60.0

50.0

40.0

30.0

20.0

10.0

0.0

NEXTflex Rapid DNA-Seq Competitor N’s Kit

50.0

45.0

40.0

35.0

30.0

25.0

20.0

15.0

10.0

5.0

0.0NEXTflex Rapid DNA-Seq Competitor N’s Kit

% Representation

Non-ligated Singly-ligated Doubly-ligated

Sample Peak 1 Peak 2 Peak 3

NEXTflex RapidDNA-Seq 10.9 42.1 47.1

Competitor N’s Kit 38.7 39.8 21.5

Figure 4. Direct comparison of NEXTflex Rapid DNA-Seq to Competitor N’s kit by analysis of proportions of ligation products. Red portions of the graph indi-cate non-ligated product pro-portion; yellow, singly-ligated; green, the desired doubly-ligated amplicon.

Figure 3. Overlays of the triplicate samples of NEXTflex Rapid DNA-Seq (left) and Competitor N’s kit (right).

Figure 5. Average triplicate values for the desired doubly-ligated products for NEXTflex Rapid DNA-Seq and Competitor N prepared libaries. The NEXT-flex libraries produced 47.1% relative conversion com-pared to Competitor N’s 21.5%.

Cat # Product Quantity

5144-01 NEXTflex™ Rapid DNA Sequencing Kit 8 rxns

5144-02 NEXTflex™ Rapid DNA Sequencing Kit 48 rxns

514120 NEXTflex™ ChIP-Seq Barcodes - 6 48 rxns




514124 NEXTflex-96™ ChIP-Seq Barcodes - 96 768 rxns

514101 NEXTflex™ DNA Barcodes - 6 48 rxns




514105 NEXTflex-96™ DNA Barcodes - 96 768 rxns

Competitive Analysis

The NEXTflex™ Rapid DNA-Seq Kit is compared below with Compet-itor N’s Kit, illustrating the advantage of Bioo Scientific’s Enhanced Adapter Ligation Technology. The NEXTflex Rapid DNA-Seq Kit and the competitor kit were used to prepare samples through ligation. All samples were then purified and size-selected using an identical AMPureXP bead cleanup step. Triplicate overlays of the three liga-tion product peaks for NEXTflex Rapid DNA-Seq and Competitor N’s kits were generated. Integration of the ligation product peaks using the Agilent 2100 Bioanalyzer software produces size-independent values for percentage of product comprising each peak. These values were averaged for the triplicate ligation reactions for each sample. The resulting comparison of unligated, singly-ligated, and doubly-ligated products wAS graphed.


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NEXTflex™ DNA Sequencing Kitsfor Illumina®

The NEXTflex™ DNA Sequencing Kit is designed to prepare single, paired-end and multiplexed genomic DNA libraries for sequencing using Illu-mina® GAIIx, HiSeq and MiSeq platforms. The NEXTflex DNA Sequencing Kit features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. With an improved enzymatic ligation mix, ligations can now be performed with longer adapters and higher binding efficiencies. This NEXTflex kit simplifies workflow by using master mixed reagents and magnetic bead based clean-up, reducing pipetting and eliminating time consuming steps in library preparation. In addition, the availability of up to 96 unique adapter bar-codes makes the NEXTflex kits the most high-throughput kits available (see page 26 for more information about the NEXTflex DNA Barcodes).

What is Enhanced Adapter Ligation Technology?

• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads• Completely gel-free library prep protocol available• Flexible barcode options - Kits contain 6, 12, 24, 48 or 96 unique barcodes• Cost-effective library prep solution• Simplified workflow with master mixes reduces hands-on time• Compatible with targeted sequence capture• Bead-based cleanup• Compatible with the Illumina® GAIIx, HiSeq and MiSeq sequencing platforms

By using ligases that are designed to repair DNA nicks at a significantly higher rate (>109 compared to 106) than traditional commercial T4 DNA ligases, Bioo Scientific has revolutionized adapter ligation in library preparation. This, along with proprietary modifications to the adapter sequences designed to reduce dimer formation and increase dual adapter ligation, comprise Enhanced Adapter Ligation Technology. Using NEXTflex ligation mixes, the user can expect a significantly greater number of unique reads, sequence diversity and coverage.

Sequencing-Ready DNA Libraries from 1 ng – 1 µgof DNA in 2 Hours or Less

Visit Page 6 of the catalog or BiooNGS.com to accelerate your library prep.

• < 2 hour library construction• Largest number of sequencing reads available• Broad input range



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GENOMIC DNA

FRAGMENT

END-REPAIR

ADD ‘A’

ADD ADAPTERS

LIGATION

GEL FREE SPRI SIZE SELECTION

BRIDGE AMPLIFICATION(CLUSTER GENERATION)

COMPLEXITY REDUCTION(SEQUENCE CAPTURE)

PCR

BEAD CLEANUP

GEL SIZE SELECTION

= N= A= T= Adapters with

Cluster Sequence

NEXTflex™ DNA Sequencing Kit Protocol


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New, Completely Gel-Free, DNA Library Prep Protocol

Bioo Scientific has updated the NEXTflex DNA Sequencing Kit to include an optional protocol which allows you to completely eliminate the gel purification step. The alternative bead-based protocol reduces the required sample prep time by two hours and allows for complete automation of DNA library prep.

Deep Sequencing of the E. coli GenomeAs can be seen in Table 1, sequencing of the E. coli genome was performed using the NEXTflex™ DNA Sequencing Kit and Barcodes and compared to Company X’s protocol. The NEXTflex™ Enhanced Adapter Ligation Technology resulted in significantly improved reads, coverage and assembly. The E. coli genome was assembled with fewer scaffolds (result of connecting contigs by linking data), had an increased average maximum scaffold size and N50.

Table 1. Summary of Sequence Data between NEXTflex™ and Competitor X’s Libraries

NEXTflex™ DNA Sequencing Kit Competitor X’s Protocol

Sample E. coli E. coli

Genome Size 5 Mb 5 Mb

Read Length 36 bp 36 bp

Insert Size (bp) 200 200

Number of Reads 29,718,673 19,311,940

Read Coverage 115 75

Perfectly Mapped Reads 96% 89%

# of Unknown Bases 3,201 6,540

Contig N50 102,408 95,299


5140-01 NEXTflex™ DNA Sequencing Kit 8 rxns






514105 NEXTflex-96™ DNA Barcodes (in 96-well plate format) 768 rxns

514106 NEXTflex-96™ DNA Barcodes (in tubes) 768 rxns


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NEXTflex™ ChIP Sequencing Kitsfor Illumina®

The NEXTflex™ ChIP-Seq Kit contains specially designed enzymes and buffers needed for library preparation of chromatin immunoprecipitated (ChIP) DNA or small genomic DNA samples for next generation sequencing. The NEXTflex ChIP-Seq Kit features Bioo Scientific’s proprietary Enhanced Adapter Ligation Technology (see page 8 for more information) resulting in library preps with a larger number of unique sequencing reads. With an improved enzymatic ligation mix, ligations can now be performed with longer adapters and higher binding efficiencies. The NEXT-flex ChIP-Seq Kit simplifies workflow by using master mixed reagents and magnetic bead based cleanup, reducing pipetting and eliminating time consuming steps in library preparation. Using NEXTflex™ ChIP-Seq Barcodes (see page 26 for more information) with Enhanced Adapter Ligation Technology, the user can index up to 96 samples at once, providing an unparalleled ChIP sequencing capacity.

• Low 1 - 10 ng input requirement• For use with ChIP or genomic DNA samples• Faster protocol - Bead-based cleanup shaves hours off of library prep• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads • 96 barcoded adapters for multiplexing contain embedded index sequence• Automation-friendly workflow is compatible with liquid handlers• Flexible barcode options – Kits contain 6, 12, 24, 48 or 96 unique barcodes• Compatible with Illumina® GAIIx, HiSeq and MiSeq sequencing platforms


5143-01 NEXTflex™ ChIP Sequencing Kit 8 rxns

5143-02 NEXTflex™ ChIP Sequencing Kit 48 rxns

514120 NEXTflex™ ChIP Sequencing Barcodes - 6 48 rxns





Sequencing-Ready DNA Libraries from 1 ng – 1 µgof DNA in 2 Hours or Less

Visit Page 6 of the catalog or BiooNGS.com to accelerate your library prep.

• < 2 hour library construction• Largest number of sequencing reads available• Broad input range• Low input requirements ideal for ChIP-Seq experiments


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NEXTflex™ ChIP Sequencing Kit Protocol

END-REPAIR

IMMUNOPRECIPITATION

ADD ‘A’

ADD ADAPTERS

LIGATION

PRE-SIZE SELECTION PCR / SIZE SELECTION

30 Minutes(Optional Stop Point)

30 Minutes


1-2 Hours(Optional Stop Point)



PCR

BEAD CLEANUP

1 Hour(Optional Stop Point)

PROTEIN


Cluster Sequence


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NEXTflex™ Methyl-Seq 1 Kitsfor Illumina®

The NEXTflex™ Methyl-Seq 1 Kit is designed to enrich and prepare single, paired-end and multiplexed methylated DNA libraries for sequencing using Illumina® MiSeq, GAIIx and HiSeq platforms. NEXTflex Methyl-Seq utilizes a versatile immune-capture (MeDIP) or methyl-CpG-binding domain (MeCAP) method for detection of methylated DNA, allowing the user to easily assess the methylation state of the genome. This kit features Enhanced Adapter Ligation Technology (see page 8 for more information), resulting in library preps with a larger number of unique sequencing reads. This specially designed NEXTflex ligation enzymatic mix allows users to perform ligations with longer adapters and bet-ter ligation efficiencies. The NEXTflex Methyl-Seq 1 Kit simplifies workflow by using master mixed reagents and magnetic bead based cleanup, reducing pipetting and eliminating time consuming steps in library preparation. In addition, the availability of the NEXTflex™ DNA Barcodes (see pg 26) with up to 96 unique adapter indices makes the NEXTflex kits the most high-throughput kits available.

Table 2: Sequence Comparison between Differentiated and Non-differentiated Cells

Figure 6. DNA Methylome, Genome Scale

Figure 6 shows that on a genome scale, DNA methylomes of human embryonic stem cells and iPSCs transformed from a human dermal fibro-blast cell are similar to one another and highly distinct from the primary somatic cell lines.

• Methylome-level assessment with broad genome coverage• Make methyl rich libraries using methylated DNA immunoprecipitation (MeDIP) or MBD capture (MeCAP)• Quantify absolute DNA methylation levels• Identify differentially methylated regions (DMRs)• Enhanced Adapter Ligation Technology with NEXTflex™ Ligation• Flexible barcode options– 6, 12, 24, 48 or 96 unique indices• Gel-free and bead-based cleanup protocols• Automation-friendly workflow is compatible with liquid handlers• Barcoded adapters for multiplexing contain embedded index sequence• Functionally validated with Illumina® MiSeq, GAIIx and HiSeq platforms

HDF-FibroblastHDFH1

HDF-iPSC

Chromosome 10: 131,130,993 - 132,030,027mC(ATC)GmC(ATC)(ATC)

Sample CulturePassage

PCRCycles

CaptureMethod Format Unique

Mapped ReadsDMRs

CoveredmCs

Covered

H1 25-27 10 MeDIP Single Read 120,154,231 87.89% 91.38%

HDF - 9 MeDIP Paired Read 79,286,174 88.93% 93.49%

HDF - 9 MeCAP Paired Read 76,884,138 89.63% 94.50%

HDF-iPSC 15 8 MeCAP Paired Read 83,290,387 83.34% 87.64%


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Figure 7. Frequency of DNA Methylation at Both CG and Non-CG sites SuggestiPSCs Resemble ES and are Distinct from Somatic Cells

Figure 7 shows that high sequence coverage of four DNA methylomes allowed interrogation of 87-94% of genomic methylated cytosines using both MeDIP and MeCAP enrichment. Pluripotent cell coverage is significantly different from somatic cell lines using both enrichment methods.

90

80

70

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50

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10

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HDF-Fibroblast

HDF

HDF-iPSC

H1Met

hyla

ted

CG/C

G (

%)


5118-01 NEXTflex™ Methyl Sequencing 1 Kit 8 rxns

5118-02 NEXTflex™ Methyl Sequencing 1 Kit 48 rxns





514105 NEXTflex-96™ DNA Barcodes - 96 (in 96-well plate format) 768 rxns

514106 NEXTflex-96™ DNA Barcodes - 96 (in tubes) 768 rxns


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NEXTflex™ Methyl-Seq 1 Kit Protocol

GENOMIC DNA

FRAGMENT

END REPAIR

ADENYLATION

LIGATION

METHYLATED DNA ENRICHMENT

PRE-SIZE SELECTION PCR

GEL SIZE SELECTION

PCR



30 Minutes

30 Minutes

15 Minutes

15 Minutes

1 Hour

1-2 Hours


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NEXTflex™ Bisulfite-Seq Kitsfor Illumina®

The NEXTflex™ Bisulfite-Seq Kit is designed to work with reduced representation or total bisulfite-converted DNA libraries and is compatible with single, paired-end and multiplexed DNA libraries on Illumina® MiSeq, GAIIx and HiSeq platforms. NEXTflex Bisulfite-Seq with reduced represen-tation utilizes a restriction enzyme that leads to selective amplification of CpG regions, resulting in focused sequence depth. The NEXTflex™ Msp1 Restriction Enzyme, which is available separately, has been optimized for reduced representation studies when used in conjunction with the NEXTflex Bisulfite-Seq Kit. The NEXTflex Bisulfite-Seq Kit can also be used without the Msp1 Restriction Enzyme for genome-wide methyl-seq analysis, including under-represented CpG regions. This kit features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. This specially designed NEXTflex ligation enzymatic mix allows users to perform ligations with longer adapters and better ligation efficiencies. The NEXTflex Bisulfite-Seq Kit is a versatile kit designed to facilitate assessment of the methylation state of the genome and simplify workflow by using master mixed reagents and magnetic bead based cleanup, reducing pipetting and eliminating time-consuming steps in library preparation. In addition, the availability of the methylated NEXTflex Bisulfite-Seq barcoded adapters make multiplexing possible.

• Compatible with total bisulfite-seq and reduced representation • Single nucleotide resolution of methylation sites• Uracil insensitive polymerase designed for bisulfite-converted DNA• Methylome-level assessment with broad genome coverage• Enhanced Adapter Ligation Technology with NEXTflex™ Ligation• Bead-based cleanup• Automation-friendly workflow is compatible with liquid handlers• Methylated barcoded adapters for multiplexing contain embedded index sequence• Functionally validated with Illumina® MiSeq, GAIIx and HiSeq platforms


5119-01 NEXTflex™ Bisulfite Sequencing Kit 8 rxns

5119-02 NEXTflex™ Bisulfite Sequencing Kit 48 rxns

511911 NEXTflex™ Bisulfite Sequencing Barcodes - 6 48 rxns


511921 NEXTflex™ Msp1 Restriction Enzyme 8 rxns

511922 NEXTflex™ Msp1 Restriction Enzyme 48 rxns


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NEXTflex™ Bisulfite-Seq Kit Protocol

END-REPAIR

ADD ‘A’

ADD METHYLATED ADAPTERS

LIGATION OF METHYLATED ADAPTERS

SIZE SELECTION

BISULFITE CONVERSION

FRAGMENT

GENOMIC DNA


30 Minutes



3 Hours



PCR

BEAD CLEANUP



Cluster Sequence


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NEXTflex™ PCR-Free DNA Sequencing Kitsfor Illumina®

The NEXTflex™ PCR-Free DNA Sequencing Kit offers all the advantages of the original NEXTflex™ DNA Sequencing Kit, in addition to completely eliminating the need for amplification. Amplifying AT or GC rich genomic regions often leads to sequence biased nucleotide compositions and poses a serious challenge during analysis. Using specially designed master-mixed enzymes, the NEXTflex PCR-Free DNA Sequencing kit completely eliminates the need for amplification, enabling better read mapping and a reduction in duplicate sequences, leading to reduced sequence cost and bias, for more representative base identities and better de novo assembly. In assessing bias, the presence of low quality and high GC reads often cannot be aligned against a reference genome. High GC-containing profiles tend to shift toward higher GC content, an indication of poor base representation. Eliminating PCR removes this content bias for more representative reads. PCR is also the cause of a significant number of gene duplicates, which increases sequence cost, adapter dimers and noise in cluster detection. Using the NEXTflex PCR-Free Kit, the number of duplicate sequences is reduced, ensuring a more representative matched number of reads. While the quantity of template generated using the NEXTflex PCR-Free DNA Sequencing Kit is lower than the NEXTflex DNA Sequencing Kit, library quantification by qPCR demonstrates that from as little as 0.5 µg of DNA, a sufficient amount of 300-500 bp PCR-free library can be obtained for greater than 600 high density HiSeq lanes. The NEXTflex™ PCR-Free Barcodes, available in sets of up to 48 barcodes, can be used to multiplex these reactions (see page 26 for more information).

• Eliminates amplification bias and poor sequence representation• Improves read mapping• Reduces duplicate sequences• Better de novo assembly• Completely gel-free protocol• Significantly faster library preparation time


5142-01 NEXTflex™ PCR-Free DNA Sequencing Kit 8 rxns

5142-02 NEXTflex™ PCR-Free DNA Sequencing Kit 48 rxns

514110 NEXTflex™ PCR-Free DNA Barcodes - 6 48 rxns





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NEXTflex™ PCR-Free DNA Sequencing Kit Protocol

GENOMIC DNA

FRAGMENT

END-REPAIR

ADD ‘A’

ADD ADAPTERS

LIGATION


BEAD CLEANUP


30 Minutes

SIZE SELECTION 1 Hour



Cluster Sequence


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NEXTflex™ Pre-Capture Combo Kit(NimbleGen SeqCap EZ Library Compatible)for Illumina®

The NEXTflex™ Pre-Capture Combo Kit (NimbleGen SeqCap EZ Library Compatible) is a complete library prep solution designed for library preparation, multiplexing and barcode blocking, upstream of Roche NimbleGen’s SeqCap EZ capture kits. The NEXTflex Pre-Capture Combo Kit (NimbleGen SeqCap Compatible) is designed to prepare single, paired-end and multiplexed genomic DNA libraries for target capture and Illu-mina® compatible sequencing. This kit features Enhanced Adapter Ligation Technology resulting in library preps with a larger number of unique sequencing reads. With Bioo Scientific’s improved enzymatic ligation mix, ligations can be performed with longer adapters and higher binding efficiencies. The NEXTflex Pre-Capture Combo Kit simplifies workflow by using master mixed reagents and magnetic bead based cleanup, reduc-ing pipetting and eliminating time consuming steps in library preparation.

Table 3. Mapping and coverage statistics for twelve sample libraries demonstrates the efficacy of pre-capture multiplexing using NEXTflex library and blocking oligo reagents.

• Complete Kit – Library prep, barcodes and barcode blockers compatible with NimbleGen’s SeqCap EZ capture systems • Simplifies and reduces the cost of target capture • Multiplex Compatible - 96 barcodes and barcode blockers available • Optimized - Enhanced Adapter Ligation Technology offers larger number of unique reads • Completely gel-free library prep protocol available • Simplified workflow with master mixes that reduces hands on time • Bead-based cleanup protocol • Compatible with the Illumina® GAIIx and HiSeq sequencing platforms


5140-51 NEXTflex™ Pre-Capture Combo Kit - 6 Barcodes 48 rxns




514131 NEXTflex™ DNA Barcode Blockers - 6 for SeqCap EZ Library 48 rxns



514134 NEXTflex-96™ DNA Barcode Blockers - 96 for SeqCap EZ Library 8 x 96 pooled libraries

Samples NEXTflex Index

Targeted Bases Raw Reads Gb of

Sequence

% Raw Reads

Mapped

% Unique Reads

Mapped

% Mapped

Reads on Target

Unique Gb of

Sequence on Target

AverageCoverage

%Sensitivity

inDetecting

Known SNPs

%Specificity in

DetectingKnown SNPs

1 CGATGT 3898910 26,548,162 4.0 90.4 78.8 63.4 2.6 220.4 97.70 98.70

2 TGACCA 3898910 25,218,056 3.8 90.3 79.2 63.6 2.4 211.1 98.10 98.90

3 ACAGTG 3898910 27,026,818 4.1 90.7 79.0 64.2 2.6 227.9 97.90 99.00

4 GCCAAT 3898910 24,589,892 3.7 90.2 78.8 63.5 2.4 204.7 97.70 98.60

5 CAGATC 3898910 26,077,566 4.0 90.5 79.3 63.9 2.5 219.4 97.80 98.70

6 CTTGTA 3898910 24,412,142 3.7 90.4 80.1 63.4 2.4 206.2 98.00 98.90

7 ATCACG 3898910 29,317,270 4.5 90.7 78.2 64.2 2.9 244.6 97.80 98.90

8 TTAGGC 3898910 20,994,732 3.2 90.5 80.0 64.0 2.0 178.6 97.70 98.40

9 ACTTGA 3898910 25,344,596 3.9 90.5 79.8 63.4 2.4 213.3 97.70 98.60

10 CATCAG 3898910 25,371,206 3.9 90.4 79.5 63.9 2.5 214.5 98.00 98.90

11 TAGCTT 3898910 27,844,310 4.2 90.6 79.3 63.9 2.7 234.6 97.80 99.00

12 GGCTAC 3898910 23,987,332 3.6 90.4 79.3 64.0 2.3 202.2 97.70 98.70


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NEXTflex™ Pre-Capture Combo Kit Protocol

GENOMIC DNA

FRAGMENT

END REPAIR

ADD ‘A’

ADD ADAPTERS

LIGATION

SIZE-SELECTION

PCR

BEAD CLEANUP

TARGET CAPTUREUsing NimbleGen SeqCap EZ Library Kit

SEQUENCING


30 Minutes





Cluster Sequence


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NEXTflex™ 16S V4 Amplicon-Seq Kitfor Illumina®

The NEXTflex™ 16S V4 Amplicon-Seq Kit is a validated amplicon-seq library prep kit developed to simplify bacterial metagenomics studies using Illumina® MiSeq and HiSeq platforms. This kit allows users to go from sample to sequence in two hours, making it the fastest amplicon-seq library prep kit available. The NEXTflex 16S V4 Amplicon-Seq Kit offers a fast two step, gel-free protocol that reduces library prep time to 2 hours and allows for complete automation of bacterial metagenomics studies. There are two main steps involved in the NEXTflex 16S V4 Amplicon-Seq Kit protocol: amplification and cleanup. Primers included in the NEXTflex 16S V4 Amplicon-Seq Kit are designed to hybridize to the fourth hypervariable (V4) domain of microbial 16S ribosomal RNA (rRNA) genes allowing for amplification and sequencing of the variable regions. The single cleanup step ensures maximum recovery of amplicons for downstream sequencing.

• Fast, two-step library prep protocol • Flexible barcode options - Kits containing 4, 12, 24, 48, 96 or 288 unique barcodes • Automation-friendly workflow is compatible with liquid handlers • Cost-effective library prep solution • Only 50 ng of input required • Functionally validated with Illumina® MiSeq and HiSeq sequencing platforms


4201-01 NEXTflex™ 16S V4 Amplicon-Seq Kit - 4 Barcodes 16 rxns





4201-07 NEXTflex™ 16S V4 Amplicon-Seq Kit - 288 Barcodes 1,152 rxns

NEXTflex™ 16S V4 Amplicon-Seq Kit Protocol

SEQUENCING

PCR

BACTERIAL GENOMIC DNA

BEAD CLEANUP

2 Hours

V4 Reverse

V4 Forward


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NEXTflex™ DNA Sequencing Kit(BioMek® FXp Compatible)for Illumina®

The NEXTflex™ DNA Sequencing Kit for Biomek® FXp is designed to prepare single, paired-end and multiplexed Illumina®-compatible genomic DNA libraries using the Biomek® FXp Span8/MC hybrid liquid handler. Minimized setup time for high-throughput sequencing using the NEXTflex DNA Sequencing Kit for Biomek FXp means less labor time and more consistent sample preparation. The NEXTflex DNA Sequencing Kit fea-tures Enhanced Adapter Ligation Technology resulting in library preps with a larger number of unique sequencing reads. With Bioo Scientific’s improved enzymatic ligation mix, ligations can be performed with longer adapters and better binding efficiencies. Output genomic DNA libraries are compatible with the Illumina® GAIIx, HiSeq and MiSeq platforms. In addition, sets of up to 96 unique adapter barcodes are available for multiplexing. The library preparation is automated using the SPRIWorks HT protocol on the Beckman Coulter Biomek FXp Span8/MC hybrid liquid handler.

Competitive AnalysisTable 4: Comparison of NEXTflex™ DNA-Seq Kit for Biomek and Competitor’s Kits

• Library preparation kit for use with the Biomek® FXp Liquid Handler• Optimized - Enhanced Adapter Ligation Technology offers larger number of unique reads• Multiplex Compatible – Sets of up to 96 barcodes are available• Cost-effective library prep solution• Automated bead-based cleanup protocol• Compatible with Illumina® GAIIx, HiSeq and MiSeq sequencing platform


5140-41 NEXTflex™ DNA Sequencing Kit (BioMek® FXp Compatible) 48 rxns





514105 NEXTflex-96™ DNA Barcodes (in 96-well plate format) 768 rxns

514106 NEXTflex-96™ DNA Barcodes (in tube format) 768 rxns

NEXTflex™ DNA Sequencing Kit for BioMek® FXp Competitor X’s Protocol

Enhanced Adapter Ligation Technology Yes No

Multiplex Barcodes 96 No

Magnetic Bead Purification Yes Yes

Compatible with SPRIWorks HT Protocol Yes Yes

Gel-Free Protocol Yes Yes

Price/Reaction Lower Higher


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NEXTflex™ DNA-Seq Modulesfor Illumina®

Bioo Scientific now offers NEXTflex™ DNA, ChIP-Seq, and PCR-Free Modules allowing even more flexibility for NGS library prep. These modules are provided in a convenient master mix format to reduce steps during DNA sample prep workflows. Modules are available for each step in the library preparation protocol. Every NEXTflex Module passes rigorous enzymatic quality control and is functionally validated by sequencing on an Illumina® platform. The NEXTflex™ Ligation modules incorporate Bioo Scientific’s proprietary Enhanced Adapter Ligation Technology, which results in library preps with a larger number of unique sequencing reads. With Bioo Scientific’s improved enzymatic ligation mix, ligations can be performed with longer adapters and better ligation efficiencies.

NEXTflex™ DNA Modules

These modules are also components of the NEXTflex™ DNA Sequencing Kit. The NEXTflex DNA Modules offer additional flexibility for next gen-eration sequencing library preparation. They can be used to prepare single, paired-end and multiplexed genomic DNA libraries for sequencing. These modules should be used with the NEXTflex™ DNA Barcodes for multiplexing (see pg 26).

NEXTflex™ ChIP-Seq Modules

The NEXTflex™ ChIP-Seq Modules offer additional flexibility for next generation sequencing library preparation. The NEXTflex ChIP-Seq Modules contain specially designed NanoQ™ enzymes and buffers needed for sample preparation of chromatin immunoprecipitated or regular genomic DNA for next generation sequencing.

• Flexible options – The NEXTflex™ Modules are available in aliquots containing any number of reactions• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads• Automation-friendly workflow is compatible with liquid handlers• Convenient – Reactions are provided in master mix format to reduce steps during DNA sample prep workflows• Functionally validated with Illumina® sequencing platforms

• 1 µg input requirement• For use with genomic DNA samples

• Low 10 ng input requirement• For use with ChIP or genomic DNA samples• End repair and ligation mixes are optimized for low DNA input


5140-05 NEXTflex™ DNA End Repair Mix 100 rxns

5140-06 NEXTflex™ DNA Adenylation Mix 100 rxns

5140-07 NEXTflex™ DNA Ligation Mix 100 rxns

5140-08 NEXTflex™ DNA PCR Master Mix 100 rxns


5143-05 NEXTflex™ ChIP Sequencing End Repair Mix 100 rxns

5143-06 NEXTflex™ ChIP Sequencing Adenylation Mix 100 rxns

5143-07 NEXTflex™ ChIP Sequencing Ligation Mix 100 rxns

5143-08 NEXTflex™ ChIP Sequencing PCR Master Mix 100 rxns


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NEXTflex™ PCR-Free Modules

These modules are also components of the NEXTflex™ PCR-Free DNA Sequencing Kit. These modules should be used with the NEXTflex™ PCR-Free Barcodes for multiplexing (see pg 26).

NEXTflex™ DNA-Seq Multiplexing Optionsfor Illumina®

The NEXTflex™ and NEXTflex-96™ DNA Barcodes are adapters containing indexed sequences that offer an improved multiplexing workflow and flexible setup. This automation-friendly format enables multiplexing of up to 96 samples. The NEXTflex DNA Barcodes are available in sets of 6, 12, 24 and 48 unique adapters. For scientists who want to multiplex more than 48 samples, the NEXTflex-96 DNA Barcodes are sets of 96 barcodes that are available in microfuge tube or 96-well plate formats. The ability to pool samples in an efficient way significantly decreases hands-on time while providing robust data quality.

Figure 8. NEXTflex Adapter Design

The NEXTflex™ adapters contain the full complement of sequencing flow cell binding regions (A, B), which eliminates the need to perform PCR to add the barcode tag.

Figure 9. Deep Sequencing of NEXTflex DNA Barcode Generated Sequencing Libraries. Libraries generated using the NEXTflex DNA Barcodes have a consistent percentage of usable reads.

• 0.5 - 3 µg input requirement• For use with genomic DNA samples

A

Read 1 DNA Insert Index

Read 2 B

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01 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47


5142-05 NEXTflex™ PCR-Free End Repair Mix 100 rxns

5142-06 NEXTflex™ PCR-Free Adenylation Mix 100 rxns

5142-07 NEXTflex™ PCR-Free Ligation Mix 100 rxns


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The NEXTflex™ Barcodes simplify high-throughput sequencing with up to 96 available adapters. Using the NEXTflex™ DNA Sequencing Kit, 48 libraries were prepared and tagged with a different barcoded adapter. The percentage of index reads that could be used is represented by the height of the bars in Figure 5. Most barcoded adapters were read perfectly. The NEXTflex™ index utilizes double error correction, ensuring that single base changes during phasing do not unrecognizably alter the barcode sequence.

Barcodes for NEXTflex™ ChIP-Seq & Rapid DNA-Seq Kits

Barcodes for NEXTflex™ Rapid DNA-Seq, DNA-Seq & Methyl-Seq 1 Kits

Barcodes for NEXTflex™ PCR-Free DNA-Seq Kits

Barcodes for NEXTflex™ Bisulfite-Seq Kits












514105 NEXTflex-96™ DNA Barcodes 768 rxns


514110 NEXTflex™ PCR-Free Barcodes - 6 48 rxns








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AIR™ DNA-Seq Multiplexing Optionsfor Illumina®

AIR™ DNA Barcodes contain 12, 24, 36 or 48 unique barcodes, which can be used to provide flexibility in high-throughput sequencing applica-tions. They significantly increase scale and throughput while reducing costs by allowing the user to pool up to 48 multiple library preparations into a single flow cell. The AIR DNA Barcodes accomplish this by using a 6 nt index on the PCR primer. This allows for proper differentiation between samples, preventing poor reads from single base errors introduced during PCR.


513901 AIR™ DNA Barcodes - 12 96 rxns




KEY FEATURES

• Pre-programmed with NEXTflex™ library prep protocols

• Fully automated, hands-off NGS library prep

• Open platform

• Scalable for processing of up to 96 samples in parallel

Imagine how much money and time you could save with your NGS library preparation.

See page 54 for more information.



DNA-Seq Library PrepIon PGM™ & Ion Proton™ Compatible


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NEXTflex™ DNA Sequencing Kitsfor Ion PGM™ & Ion Proton™

The NEXTflex™ DNA Sequencing Kit for Ion PGM™ and Ion Proton™ is designed to prepare genomic DNA or ChIP-Seq libraries for sequencing using Life Technologies’ Ion PGM or Ion Proton sequencing platform. The NEXTflex DNA Sequencing Kit features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. This NEXTflex Kit simplifies workflow by using master mixed reagents and bead-based cleanup. In addition, the ability to perform the reactions in a 96-well plate makes this kit more automation-friendly than other commercially available kits.

Competitive Analysis Table 5: Comparison between NEXTflex™ and Competitor’s DNA Library Prep Kits for the Ion PGM™ & Ion Proton™

NEXTflex™ DNA-Seq Multiplexing Optionsfor Ion PGM™ & Ion Proton™

The NEXTflex™ DNA Barcodes for Ion PGM™ and Ion Proton™ are adapters containing indexed sequences that offer an improved multiplexing workflow and flexible setup. This format enables multiplexing of up to 64 samples. The NEXTflex DNA Barcodes are available in sets of 8, 16, 32 and 64 unique adapters. The ability to pool samples in an efficient way significantly decreases hands-on time while providing robust data quality.

• Flexible barcode options - Kits contain 8, 16, 32 or 64 unique barcodes• Simplified workflow with 96-well plate compatible volumes• Compatible with DNA-Seq & ChIP-Seq• Compatible with Life Technologies’ Ion Torrent sequencing platform










96-Well Plate Compatible Yes No

Automation Friendly Yes Less So

Size Selection Components Included Yes No

Starting Material Required 10 ng - 1 µg 10 ng - 1 µg


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GENOMIC DNA

FRAGMENT

END REPAIR 30 Minutes(Optional stop point)

15 Minutes(Optional stop point)

1-2 Hours(Optional stop point)

1 Hour(Optional stop point)

ADD ADAPTERSP1 Adapter

P1 Adapter

A AdapterCTGAGACT

LIGATIONA Adapter

CTGAGACT

P1 Adapter

SIZE-SELECTIONA Adapter

CTGAGACT

P1 Adapter

PCR

DILUTION FACTOR DETERMINATIONBIOANALYZER AND/OR qPCR

DNA TEMPLATE PREPARATION

A Adapter

CTGAGACT

GACT

Key Sequence



DNA-Seq Library PrepSOLiD Compatible


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NEXTflex™ DNA Sequencing Kitsfor 5500 SOLiD™

The NEXTflex™ DNA Sequencing Kit for 5500 SOLiD™ is designed to prepare single, paired-end and multiplexed genomic DNA or ChIP-Seq libraries for sequencing using Life Technologies’ 5500 SOLiD sequencing platform. The NEXTflex DNA Sequencing Kit for 5500 SOLiD features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. This NEXTflex kit simplifies workflow by using master mixed reagents and bead-based cleanup. In addition, the ability to perform the reactions in a 96-well plate makes this kit more automation-friendly than other commercially available kits.

Competitive Analysis Table 6: Comparison between NEXTflex™ and Competitor’s DNA Library Prep Kits

NEXTflex™ DNA Barcodesfor 5500 SOLiD™

The NEXTflex™ DNA Barcodes for 5500 SOLiD™ are adapters containing indexed sequences that offer an improved multiplexing workflow and flexible setup. This format enables multiplexing of up to 96 samples. The NEXTflex DNA Barcodes for 5500 SOLiD sets A - E contain 8 rxns each of 16 unique barcodes; set F contains 8 rxns each of all 96 barcodes.

• Simplified workflow with 96-well plate compatible volumes• Flexible barcode options - Barcode kits contain 16 to 96 unique barcodes• Simplified workflow with master mixes reduces hands-on time• Compatible with ChIP-Seq


4101-01 NEXTflex™ DNA Sequencing Kit for 5500 SOLiD 12 rxns

4101-02 NEXTflex™ DNA Sequencing Kit for 5500 SOLiD 48 rxns

411001 NEXTflex™ DNA Barcodes Set A for 5500 SOLiD 128 rxns

411002 NEXTflex™ DNA Barcodes Set B for 5500 SOLiD 128 rxns

411003 NEXTflex™ DNA Barcodes Set C for 5500 SOLiD 128 rxns

411004 NEXTflex™ DNA Barcodes Set D for 5500 SOLiD 128 rxns

411005 NEXTflex™ DNA Barcodes Set E for 5500 SOLiD 128 rxns

411006 NEXTflex™ DNA Barcodes Set F for 5500 SOLiD 128 rxns

411007 NEXTflex™ DNA Barcodes Set G for 5500 SOLiD 768 rxns





Size Material Required 10 ng - 1 µg 10 ng - 1 µg


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GENOMIC DNA

FRAGMENT

END REPAIR 30 Minutes

30 Minutes

BEAD SIZE-SELECTION 30 Minutes(Optional stop point)


1 Hour(Optional stop point)

ADD “A”

ADD ADAPTERSP1 Adapter Standard Adapter

P1 Adapter Standard Adapter

LIGATION

NICK TRANSLATION & PCR

DNA QUANTIFICATION & SIZE DISTRIBUTION(qPCR & BIOANALYZER)

EMULSION PCR

PCR Primer 2

PCR Primer 1


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NEXTflex™ DNA Sequencing Kitsfor SOLiD™ 4

The NEXTflex™ DNA Sequencing Kit for SOLiD™ 4 is designed to prepare single, paired-end and multiplexed genomic DNA or ChIP-Seq librar-ies for sequencing using Life Technologies’ SOLiD 4 sequencing platform. The NEXTflex DNA Sequencing Kit for SOLiD 4 features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. This NEXTflex kit simplifies workflow by using master mixed reagents and gel-based cleanup. In addition, the ability to perform the reactions in a 96-well plate makes this kit more automation-friendly than other commercially available kits.

Competitive Analysis Table 7: Comparison between NEXTflex™ and Competitor’s DNA Library Prep Kits

NEXTflex™ DNA Barcodesfor SOLiD™ 4

The NEXTflex™ DNA Barcodes for SOLiD™ 4 are adapters containing indexed sequences that offer an improved multiplexing workflow and flex-ible setup. This format enables multiplexing of up to 96 samples. The NEXTflex DNA Barcodes for SOLiD 4 sets A - E contain 8 rxns each of 16 unique barcodes; set F contains 8 rxns each of all 96 barcodes.

• Simplified workflow with 96-well plate compatible volumes• Flexible barcode options - Barcode kits contain 16 to 96 unique barcodes• Simplified workflow with master mixes reduces hands-on time• Compatible with ChIP-Seq


4102-01 NEXTflex™ DNA Sequencing Kit for SOLiD 4 12 rxns

4102-02 NEXTflex™ DNA Sequencing Kit for SOLiD 4 48 rxns

411101 NEXTflex™ DNA Barcodes Set A for SOLiD 4 128 rxns

411102 NEXTflex™ DNA Barcodes Set B for SOLiD 4 128 rxns

411103 NEXTflex™ DNA Barcodes Set C for SOLiD 4 128 rxns

411104 NEXTflex™ DNA Barcodes Set D for SOLiD 4 128 rxns

411105 NEXTflex™ DNA Barcodes Set E for SOLiD 4 128 rxns

411106 NEXTflex™ DNA Barcodes Set F for SOLiD 4 128 rxns

411107 NEXTflex™ DNA Barcodes Set G for SOLiD 4 768 rxns





Size Material Required 10 ng - 1 µg 10 ng - 1 µg


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GENOMIC DNA

FRAGMENT

END REPAIR 30 Minutes

BEAD SIZE-SELECTION 30 Minutes(Optional stop point)


ADD ADAPTERSP1 Adapter P2 Adapter

P1 Adapter P2 Adapter

LIGATION

NICK TRANSLATION & PCR

DNA QUANTIFICATION & SIZE DISTRIBUTION(qPCR & BIOANALYZER)

EMULSION PCR

PCR Primer 2

PCR Primer 1



RNA-Seq Library PrepIllumina® Compatible


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• Enables high precision measurement of RNA concentration• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads• Input 10 - 100 ng of mRNA or rRNA-depleted total RNA• Embedded sample index barcodes for multiplexing of up to 96 samples• Automation-friendly workflow is compatible with liquid handlers• Functionally validated with Illumina® sequencing platforms

NEXTflex™ qRNA-Seq™ Kitfor Construction of Molecular Indexed ™ Libraries

Bioo Scientific’s patent pending NEXTflex™ qRNA-Seq™ Kit, enables high precision gene expression analysis by RNA-Seq. Developed in con-junction with Cellular Research Inc.™, this new kit efficiently generates libraries equivalent to conventional RNA-Seq libraries, but with the added feature of molecular indexing. Similar to conventional RNA-Seq, sample RNA is converted to cDNA fragments. Prior to PCR amplification, all cDNA fragment ends are ligated to pairs of adapters chosen at random from a total set of 9,216 molecular indices. Individual DNA molecules of identical sequence become distinct through indexing (Figure 10), allowing for differentiation between re-sampling of the same molecule and sampling of a different molecule of identical sequence. Analysis using molecular indexing information provides an absolute, digital measure-ment of gene expression levels, irrespective of common amplification distortions observed in many RNA-Seq experiments.

Figure 10. An illustration with 8 mapped reads to an mRNA transcript either with or without molecular indexing. Individual molecular indices at fragment ends are represented by different colors. The number of cDNA fragments represented by the reads is unknown without molecular indexing, but can be determined with molecular indexing.

Stochastic Labeling of Individual DNA MoleculesThe NEXTflex qRNA-Seq Kit contains two sets of 96 distinct molecular labels on the sequencing adapters. Each label or index consists of an 8 nucleotide barcode tag. In the ligation reaction, these 96 adapters are present in vast molar excess over the concentration of the cDNA fragments, and therefore serve as a non-depleting reservoir of molecular labels. Each end of a cDNA fragment independently and randomly chooses and ligates to a single label from this pool of 96 adapters to result in a total of 96 x 96 = 9,216 possible combinations across both ends. For every clone sequenced, paired-end reads reveal the chosen label on each end along with adjoining cDNA sequence. In addition to encoding DNA fragments at the molecular level, the kit also allows for the application of sample-specific barcodes during the library preparation PCR step. A more detailed description of the use of molecular indexing for RNA-Seq is available in our product application note.

ProtocolUsing the NEXTflex qRNA-Seq Kit, mRNA or rRNA-depleted RNA is fragmented using a cationic buffer. Fragmented RNA undergoes first and second strand synthesis, followed by end-repair, adenylation, ligation and PCR.

mRNA

Conventional RNA-Seq Without Molecular Indexing

8 reads Unknown # of fragments

8 reads 4 fragments

8 reads 6 fragments

8 reads 8 fragments

NEXTflex™ qRNA-Seq™ With Molecular Indexing


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Multiplexing NEXTflex qRNA-Seq Kits include barcodes. The 8 rxn kit includes 4 barcodes for multiplexing. Four different versions of the 48 reaction kit are available, each containing 24 unique barcodes. Up to 96 samples can be multiplexed at once.

POLY A SELECTED RNA

FRAGMENTATION

FIRST STRAND cDNA SYNTHESIS

SECOND STRAND SYNTHESIS

END REPAIR

ADENYLATION

ADD MOLECULAR ENCODING ADAPTERS

ADAPTER LIGATION AND LIBRARY ENRICHMENT PCR

CLUSTER GENERATION AND SEQUENCINGRead 1 Sample 1

Read 2


5130-01 NEXTflex™ qRNA-Seq™ Kit - 4 8 rxns

5130-02 NEXTflex™ qRNA-Seq™ Kit - 24 - Set A 48 rxns

5130-03 NEXTflex™ qRNA-Seq™ Kit - 24 - Set B 48 rxns

5130-04 NEXTflex™ qRNA-Seq™ Kit - 24 - Set C 48 rxns

5130-05 NEXTflex™ qRNA-Seq™ Kit - 24 - Set D 48 rxns


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• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads• Input 10 - 100 ng of mRNA or rRNA depleted total RNA• Up to 96 barcodes available for multiplexing contain embedded index sequences• Automation-friendly workflow is compatible with liquid handlers• Functionally validated with GAIIx, HiSeq and MiSeq platforms

NEXTflex™ RNA Sequencing Kitsfor Illumina®

The patent pending NEXTflex™ RNA-Seq Kits provide an easy and flexible solution for generating single, paired-end and multiplexed libraries from mRNA or rRNA depleted total RNA. Using the NEXTflex RNA-Seq Kits, mRNA or rRNA depleted RNA is fragmented using a cationic buffer. Fragmented RNA undergoes first and second strand synthesis, followed by end-repair, adenylation, ligation and PCR. In addition, the availability of up to 96 unique adapter barcodes makes this the most high-throughput kit available for RNA-Seq.

Competitive Analysis Table 8: Comparison between NEXTflex™ and Competitor’s RNA Library Prep Kits



Multiplex Barcodes Available Up to 96 unique 96 dual index

Automation Friendly Yes Yes


Figure 11. Relative Levels of ERCC Control RNAs in RNA-Seq Libraries. Made from NEXTflex Poly(A) Bead Selected and Total RNA Correlation between expected and observed values for percentage of reads mapping to 94 ERCC control RNAs in RNA-Seq library made using 20 µL of NEXTflex Poly(A) Beads.

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5129-01 NEXTflex™ RNA Sequencing Kit 8 rxns

5129-02 NEXTflex™ RNA Sequencing Kit 48 rxns

512911 NEXTflex™ RNA Sequencing Barcodes - 6 48 rxns




512915 NEXTflex-96™ RNA Sequencing Barcodes (in 96-well plate format) 768 rxns

Figure 12. Relative Levels of Different Categories of Endogenous Transcripts Detected in RNA-Seq Libraries made from NEXTflex Poly(A) Bead Selected and Total RNA. Percentage of reads mapping to rRNA regions and regions of interest with and without Poly(A) bead selection.

100%

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Total RNA(A549)

Total RNA(A549)

Total RNA(293)

Total RNA(293)

Bioo Beads20 µL

Bioo Beads50 µL

Bioo Beads100 µL

unaligned

intergenic

intron

exon

misc_RNA

lincRNA

miRNA

Mito_gDNA_rRNA_tRNA

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NEXTflex™ RNA Sequencing Kit Protocol

END-REPAIR

ADD ‘A’

ADD ADAPTERS

LIGATION

PCR

BEAD CLEANUP


FIRST STRAND SYNTHESIS

POLY (A) OR rRNA DEPLETED RNA

FRAGMENTATION


75 Minutes

10 Minutes


30 Minutes




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• Offers measurement of mRNA strand orientation • dUTP-based stranded version of the NEXTflex RNA-Seq Kit • Detection of antisense transcription • Combined steps for less sample loss • Allows for single read or paired end transcriptome sequencing • Input 10 - 100 ng of mRNA or rRNA depleted total RNA • Up to 96 barcodes available for multiplexing contain embedded index sequence • Automation-friendly workflow is compatible with liquid handlers • Functionally validated with GAIIx, HiSeq and MiSeq platforms• 2 hours faster than the Directional RNA-Seq Kit (dUTP-Based)

NEXTflex™ Directional RNA-Seq Kits (dUTP-Based) v2for Illumina®

The NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) v2 provides an easy and flexible solution for generating single read, paired-end and multiplexed libraries from mRNA or rRNA depleted total RNA. The NEXTflex Directional RNA-Seq Kit (dUTP-based) v2 can be used to obtain “stranded” information which identifies the specific DNA strands a given RNA transcript was derived from. This stranded information improves transcript annotation and improves read alignment, reducing per sample sequencing costs. dUTP-based strand orientation also enables the detection of antisense expression regulatory relationships. This kit was designed to be used in conjunction with the NEXTflex™ RNA-Seq Barcodes or NEXTflex-96™ RNA-Seq Barcodes for multiplexing. The availability of up to 96 unique adapter barcodes makes this the most high-throughput kit available for RNA-seq and also allows for both single-read and paired end sequencing.

Magnetic Beads for mRNA Purification The NEXTflex™ Poly(A) Beads (pg 47) now provide a convenient method for batch purification of pure, intact mRNA upstream of NEXTflex Directional RNA-Seq library preparation. NEXTflex Poly(A) Beads beads use oligo(dT) primer to isolate polyadenylated messenger RNAs from 1 µg – 10 µg of previously isolated total RNA.


5129-07 NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) v2 8 rxns

5129-08 NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) v2 48 rxns





512915 NEXTflex-96™ RNA Sequencing Barcodes (in 96-well plate format) 768 rxns

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Figure 13. 5’ to 3’ Sequence Coverage. Normalized coverage by position. For each library, the average coverage is shown at each relative position along the transcripts’ length.


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NEXTflex™ Directional RNA-Seq Kits (dUTP-Based)for Illumina®The patent pending NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) provides an easy and flexible solution for generating single, paired-end and multiplexed libraries from mRNA or rRNA depleted total RNA. This kit is compatible with the NEXTflex™ RNA-Seq Barcodes and the NEXT-flex-96™ RNA-Seq Barcodes (pg 51). This version will be phased out at the end of 2013, replaced with the NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) v2 (pg 45).


5129-05 NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) 8 rxns

5129-06 NEXTflex™ Directional RNA-Seq Kit (dUTP-Based) 48 rxns

NEXTflex™ Directional RNA-Seq (dUTP-Based) v2 Protocol

ADD ‘A’

ADD ADAPTERS

LIGATION

UDG TREATMENT & PCR

BEAD CLEANUP


FIRST STRAND SYNTHESIS

POLY (A) OR rRNA DEPLETED RNA

FRAGMENTATION

75 Minutes

10 Minutes


30 Minutes




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• Flexible options – The NEXTflex™ Modules are available in aliquots containing any number of reactions you need• Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads• Automation-friendly workflow is compatible with liquid handlers• Convenient – Reactions are provided in master mix format to reduce steps during RNA sample prep workflows• Functionally validated with Illumina® sequencing platforms

• 0.1 - 10 µg total starting RNA• Conserve resources by selecting for mRNA prior to library prep• Magnetic bead-based protocol for easier wash step and higher recovery• No organic solvents or precipitation step required• Validated for NGS library preparation


5129-31 NEXTflex™ RNA Fragmentation Mix 100 rxns

5129-32 NEXTflex™ RNA-Seq First Strand Synthesis Mix 100 rxns

5129-33 NEXTflex™ RNA-Seq Second Strand Synthesis Mix 100 rxns

5129-34 NEXTflex™ RNA-Seq End Repair Mix 100 rxns

5129-35 NEXTflex™ RNA-Seq Adenylation Mix 100 rxns

5129-36 NEXTflex™ RNA-Seq Ligation Mix 100 rxns

5129-37 NEXTflex™ RNA-Seq PCR Mix 100 rxns

5129-42 NEXTflex™ Directional First Strand Synthesis Mix 100 rxns

5129-43 NEXTflex™ Directional Second Strand Synthesis Mix 100 rxns

5129-48 NEXTflex™ RNA-Seq UDG Mix 100 rxns


512979 NEXTflex™ Poly(A) Beads 8 rxns



NEXTflex™ RNA-Seq & Directional RNA-Seq Modulesfor Illumina®

Bioo Scientific now offers NEXTflex™ RNA-Seq Modules, offering even more flexibility to NGS library prep. These modules are provided in a convenient master mix format to reduce steps during DNA sample prep workflows. Modules are available for each step in the library prepara-tion protocol.

NEXTflex™ Poly(A) Beads

The NEXTflex™ Poly(A) Beads provide a convenient method for batch purification of pure, intact mRNA upstream of library preparation for RNA-Seq. NEXTflex Poly(A) Beads beads use oligo(dT) primer to isolate polyadenylated messenger RNAs from 0.1 µg – 10 µg of previously isolated total RNA. Separation using a magnetic stand allows for high mRNA recovery. The intact mRNA is eluted in small volumes, eliminating the need for precipitation. Ease of use and consistent results make the NEXTflex™ Poly(A) Beads ideal for NGS library preparation applications.

These beads have been validated with:

NEXTflex™ RNA-Seq Kits & NEXTflex™ RNA-Seq ModulesNEXTflex™ Directional RNA-Seq Kits & NEXTflex™ Directional RNA-Seq ModulesNEXTflex™ qRNA-Seq™ Kits


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• Eliminate ligation associated bias• Flexible barcode options with 48 barcodes available for multiplexing• Utilizes AIR™ Ligase, a highly efficient truncated T4 RNA Ligase for greater sequencing depth• Simplified workflow reduces hands-on time• Automation-friendly workflow is compatible with liquid handlers• Functionally validated with the Illumina® GAIIx, HiSeq, and MiSeq sequencing platforms

NEXTflex™ Small RNA Sequencing Kitsfor Illumina®

The NEXTflex™ Small RNA Sequencing Kits provide an easy, flexible, cost-effective solution for generating single and multiplexed libraries from small RNA using Illumina® GAIIx, HiSeq, and MiSeq sequencing platforms. This kit contains a specialized 3’ adenylated adapter that specifi-cally ligates onto microRNAs and other small RNAs containing a 3’ hydroxyl group. After adapter ligation and reverse transcription, libraries are amplified using a barcoded primer, allowing up to 48 samples to be multiplexed and sequence variations interrogated. The NEXTflex Small RNA Sequencing Kit incorporates AIR™ Ligase, an enhanced, truncated T4 RNA Ligase, which increases the efficiency with which small RNAs are tagged with adapter, giving greater sequence depth. The NEXTflex Small RNA Sequencing Kit contains all the material to prepare 24 or 48 single-plex libraries. In combination with the NEXTflex™ Small RNA Barcodes (see page 51 for more information), 48 samples can be profiled and studied simultaneously. In addition to small RNA sequencing, the kit is compatible with RIP-Seq and CLIP-Seq protocols.

Competitive Analysis Table 10: Comparison between NEXTflex™ and Competitor’s Small RNA-Seq Library Prep Kits

Eliminate Ligation Associated Bias Figure 14. Correlation of miRNA sequencing and microarray data using randomized adapter ligation technology (a) and using a conventional competitor’s small RNA library preparation procedure (b).

Bioo Scientific’s patent pending, randomized adapters allow for bias-free ligation, allowing the user to determine true representation and expression profiles of small RNAs.

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NEXTflex™ Small RNA-Seq Kit Competitor X’s Protocol

Length of Protocol Very Short Long

AIR™ Ligase Yes No

Automation Friendly Yes No

All Enzymes Included Yes No


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Figure 15. Mouse miRNA Lung Expression Levels Figure 16. Mouse miRNA Heart & Muscle Expression Levels

Citations: Jayaprakash, A. D., Jabado O., Brown, B. D. and Sachidanandam, R. (Sept 2, 2011), Identification and remediation of biases in the activity of RNA ligases in small-RNA deep sequencing Nuc Acid Res, 1–12. doi:10.1093/nar/gkr693 Nakashe, P. et al. (2011) Adapter Dimer Reduction in High-Throughput microRNA Profiling. OMICS 1(1): 6-11.

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Bioo Scientific Results mmu-mir-143

mmu-mir-1-2

mmu-mir-1-1

mmu-mir-133b

mmu-mir-133a-2

mmu-mir-133a-1

mmu-mir-451

mmu-mir-30

mmu-mir-378

0 0.1 0.2 0.3 0.4

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mmu-mir-143

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mmu-mir-133b

mmu-mir-30a

mmu-mir-1-1

mmu-mir-378

0 0.05 0.1 0.15 0.2 0.25 0.3

Muscle

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5132-01 NEXTflex™ Small RNA Sequencing Kit 24 rxns

5132-02 NEXTflex™ Small RNA Sequencing Kit 48 rxns

513301 NEXTflex™ Small RNA Barcodes (Set A) 96 rxns

513302 NEXTflex™ Small RNA Barcodes (Set B) 96 rxns

513303 NEXTflex™ Small RNA Barcodes (Set C) 96 rxns

513304 NEXTflex™ Small RNA Barcodes (Set D) 96 rxns


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NEXTflex™ Small RNA Sequencing Protocol

3’ LIGATION

5’ LIGATION (P)

REVERSE TRANSCRIPTION

FIRST PCR & SIZE SELECTION

PCR Product

3’ Adenylated AdapterRNA

3’ Adenylated AdapterLigated RNA

RT primer

Barcode Primer 1

Universal Primer

5’ Adapter

1st Strand Synthesis Product

1st Strand Synthesis Product

5’ and 3’ Adapter Ligated RNA

SECOND PCR & SIZE SELECTION

Final PCR Product

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Primer 1

Size-selected PCR Product

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RNA-Seq Multiplexing Optionsfor Illumina®

The NEXTflex™ RNA-Seq Barcodes and NEXTflex-96™ RNA-Seq Barcodes are automation-friendly adapters that are designed for use with the NEXTflex RNA-Seq Kit (see page 42) and the NEXTflex Directional RNA-Seq Kit (dUTP-Based) v2 (see page 45). These adapters allow multiplex-ing of up to 96 unique samples in a single run for higher throughput. The ability to pool samples significantly decreases hands on time while preserving robust data quality. The NEXTflex RNA-Seq Barcodes are available in sets of 6, 12, 24 and 48 unique adapters. The NEXTflex-96 RNA-Seq Barcodes are sets of 96 barcodes that are available in microfuge tube or 96-well plate formats.

Barcodes for NEXTflex™ Small RNA-Seq KitsFour sets of 12 NEXTflex™ Small RNA Barcodes with embedded index sequences are available, offering an improved multiplexing workflow and increased flexibility. These barcodes are designed for use with the NEXTflex™ Small RNA Sequencing Kit (see page 48). The ability to pool samples in an efficient way significantly decreases hands-on time while providing robust data quality. The primer barcoding system utilizes a 6 nt index to differentiate up to 48 different samples on a single flow cell lane. 48 barcoded primers are divided into 4 different kits (A-D).


512911 NEXTflex™ RNA-Seq Barcodes - 6 48 rxns




512915 NEXTflex-96™ RNA-Seq Barcodes 768 rxns


513301 NEXTflex™ Small RNA Barcodes (Set A) 96 rxns

513302 NEXTflex™ Small RNA Barcodes (Set B) 96 rxns

513303 NEXTflex™ Small RNA Barcodes (Set C) 96 rxns

513304 NEXTflex™ Small RNA Barcodes (Set D) 96 rxns

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Next-Gen Sequencing Automation


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SEQBOT™ Liquid HandlerBioo Scientific’s SEQBOT™ is a new liquid handling platform featuring an 8-channel pipette head, designed to perform up to 96 reactions per run. Fully supported recipes for Bioo Scientific kits are available on this platform, allowing the automation of these kits with no programming required.

A variety of configurations are available to meet your specific needs and budget. The base kit provides an 8-channel liquid handler with pipette mixing. This kit includes a full personal computer running Windows 7 64-bit OS, contains all programs necessary for running Bioo Scientific-provided recipes, and allows development of custom programs. For increased throughput, several additional accessories are available, including a shaker nest for full-plate shaking, an automated magnetic stand to eliminate manual transfer of the plate on and off the magnet, and a plate crane for full walk-away automation.

SEQBOT Accessory Guide

For purchasing information please call or email at [email protected]

Item Benefits Compatibility

SEQBOT Basic liquid handler All NEXTflex Kits

Plexiglas Enclosure Protects samples from environmental contamination All NEXTflex Kits

Plate Shaker Replaces pipette mixing with 96-well simultaneous mixing, increases speed All NEXTflex Kits

AutomatedMagnetic Stand

Reduces labor, eliminates manual movement of library plate on/offmagnetic stand All NEXTflex Kits

Heated Plate Shaker

Replaces pipette mixing with 96-well simultaneous mixing, increases speed, performs elevated temperature incubations

NEXTflex™ Rapid DNA-Seq Kit

Heated/CooledPlate Shaker

Replaces pipette mixing with 96-well simultaneous mixing, increases speed, performs elevated temperature incubations, performs reduced

temperature incubations

NEXTflex™ Rapid DNA-Seq KitNEXTflex™ RNA-Seq Kit

NEXTflex™ Directional RNA-Seq KitNEXTflex™ qRNA-Seq Kit

Crane Full walk-away automation possible, reduces labor by automaticallyreplacing tip boxes and moving plates

All NEXTflex Kits


Equipm

ent

Automation for Other Liquid HandlersRecipe files are also available for automation of Bioo Scientific’s most popular kits using additional liquid handler platforms, including the Beckman Coulter Biomek FX/FXp, Eppendorf epMotion, and Tecan evoFreedom. Recipe files can be customized for your preferred deck layout and available accessories. If a recipe has not yet been developed for your kit on your platform, Bioo Scientific will work with you to develop a suitable solution. Simply email us at [email protected] for your liquid handler needs.

KEY FEATURES

• Pre-programmed with NEXTflex™ library prep protocols

• Fully automated, hands-off NGS library prep

• Open platform

• Scalable for processing of up to 96 samples in parallel

Imagine how much money and time you could save with your NGS library preparation.



DNA Fragmentation & Nucleic Acid Isolationfor Next-Gen Sequencing


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• Optimized to physically shear genomic DNA into uniform fragments of genomic DNA• Offers 30% greater DNA recovery than competitor’s fragmentation protocol• Includes complete protocol and proprietary fragmentation buffer

AIR™ DNA Fragmentation Kit

The AIR™ DNA Fragmentation Kit offers a cost-effective procedure for the fragmentation of genomic DNA for next-generation sequencing sample prep. It has been optimized to physically shear genomic DNA to produce uniform fragments. The AIR DNA Fragmentation Kit shears and recovers DNA more efficiently than traditional nebulization protocols allowing for smaller sample sizes. This kit contains nebulizers, a proprietary fragmentation buffer and complete instructions. It is designed to be used with DNA sample prep protocols for Illumina®, Ion Torrent, SOLiD and Roche 454 sequencing. The procedure requires a compressed air source with a pressure regulator. Please see manual for complete details.

Figure 17. Coverage Analysis of DNA Sheared using the AIR™ DNA Fragmentation Kit. Using the AIR DNA Fragmentation kit, 1µg of Pseudomonas aeruginosa genomic DNA was fragmented, paired end libraries were constructed, and sequence data generated using one lane of an Illumina® GAIIx (2x51). There was an average of 159 reads per position and 92% of the reads mapped perfectly to Pseudomonas aeruginosa.

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5135-01 AIR™ DNA Fragmentation Kit 10 rxns

5135-02 AIR™ DNA Fragmentation Kit 40 rxns


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• Designed for NGS applications• Only six pipetting steps required• Fast and easy DNA isolation from whole blood or buffy coat/WBCs• No protease digestion step• No organic extractions• Validated with NEXTflex™ Sequencing Kits

NextPrep™ Blood DNA Isolation Kit

The NextPrep™-Blood DNA Isolation Kit is designed for the rapid purification of genomic DNA from whole blood or buffy coat for use in next generation sequencing library preparation.

Figure 18. Rapid 15 Minute DNA Isolation Protocol

DNA is recovered by solid phase extraction onto a silica matrix in a spin basket format, using a novel rapid protocol that results in extremely clean DNA. The entire procedure can be completed in ~ 15 minutes. Yields are usually several micrograms (2.5 µg – 5 µg) of high molecular weight DNA with Abs 260/280 values between 1.8 – 2.0 from a starting volume of 0.3 mL of whole blood. Yields can be increased to ~10 – 25 µg by using the modified protocol for buffy coat. The DNA is ideal for use as input for creating libraries for next generation sequencing. For this application we recommend Bioo Scientific’s NEXTflex™ DNA-Seq Kits. The recommended anticoagulant is EDTA.

Figure 19. DNA-Seq Library From Made Human Blood Using the NEXTprep™ -Blood DNA Isolation Kit

DNA isolated from HemaCare™ Donor 5C NEXTflex™ DNA Sequencing Library DNA was fragmented by nebu-lization using AIR™ Fragmentation Kit.

Fragmented DNA size-selected for 400-500 bp frag-ments on 1.2% TAE agarose gel.

Input DNA amount: 235.5 ng

Recovered: 30 µL of PCR product with a concentration of 1.08 ng/µL; 30.24 ng total

Lyse Centrifuge Dry EluteWash x 2

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3810-01 NEXTprep™ Blood DNA Isolation Kit 8 rxns

3810-02 NEXTprep™ Blood DNA Isolation Kit 48 rxns


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• Offers high yields of total RNA from tissues and cells• Includes protocol for enrichment of microRNA• Works well for recovering RNA from cell-free fluids• Includes inert co-precipitant for maximizing RNA recovery• Liquid nitrogen not required• More cost-effective than similar reagents

BiooPure™ RNA Isolation Reagent

The BiooPure™ RNA Isolation Reagent is a single-phase reagent for extraction of total RNA or enriched small RNA (including miRNA) from solid tissues and cultured cells for use in next generation sequencing library preparation. The BiooPure reagent contains guanidinium, a powerful chaotropic agent effective for rapidly inactivating nucleases, and phenol, an organic solvent used to denature and separate proteins and DNA from RNA.

Figure 20. Bioanalyzer trace of total RNA isolation using BiooPure™

The bioanalyzer trace shows data using the RNA 6000 Pico Chip Total RNA Assay. The sample was purified from A549 human lung cancer cell line. The sample was at a concentration of 4-5 ng/µL before dilution to fall in the quantitative range of the chip. The RIN number for this sample is 8.8.

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5301-01 BiooPure™ RNA Isolation Reagent 30 mL

5301-04 BiooPure™ RNA Isolation Reagent 90 mL


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• Ideal for enrichment of microRNAs and other small RNAs from eukaryotic cells and tissues• Uses spin column format• Fast and easy small RNA extraction• Offers high yields of small RNA• Validated with NEXTflex™ Sequencing Kits

• High yields of small RNA <~ 100 bases• Complete kit with all reagents except isopropanol• Linear polyacrylamide co-precipitant included

NEXTprep™ Small RNA Isolation Kit

The NEXTprep™ Small RNA Isolation Kit is designed for enrichment of small RNA from eukaryotic cells and tissues. The small RNA recovered has been verified as suitable for making small RNA libraries for next generation sequencing (NGS) using the NEXTflex™ Small RNA Sequencing Kit. The NEXTprep Small RNA Isolation Kit includes BiooPure™ RNA Extraction Reagent for initial disruption of tissues or cells. BiooPure is a single-phase reagent that contains guanidinium thiocyanate, a powerful chaotropic agent effective for rapidly inactivating nucleases, and phenol, an organic solvent used to denature and separate proteins and DNA from RNA. The kit also includes silica filter spin basket assemblies and tubes for final purification of small RNA by solid phase extraction. The kit allows recovery of small RNAs approximately 100 bases and smaller, with less than ~5% by mass of contamination with larger RNAs.

Solid tissues or cells are disrupted by grinding or vortexing in BiooPure RNA Extraction Reagent. The resulting lysate is mixed with 1-bromo-3-chloropropane and centrifuged, which results in phase separation with the RNA partitioned in the upper aqueous phase. The aqueous phase is recovered and mixed with ethanol suitable for selective precipitation of large RNA, which is removed by solid phase extraction on a silica filter. The flow-through containing small RNA is recovered and mixed with additional ethanol to precipitate the small RNA, which is then purified by solid phase extraction onto a second silica filter. The filter is washed and RNAs <~100 bases are eluted in RNase-free water containing a trace amount of EDTA.

MiraZol™ Small RNA Isolation Kit

The MiraZol™ Small RNA Isolation Kit is similar to the NextPrep™ Small RNA Isolation Kit, but uses isopropanol precipitation in place of silica filter spin columns for purification of small RNA from cells and tissue.


3812-01 NEXTprep™ Small RNA Isolation Kit 10 rxns


3812-01 NEXTprep™ Small RNA Isolation Kit 10 rxns


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Ordering Information

To place an order in the USA or Canada, please use any of the following convenient options:

To place an order in any other country, please contact your local distributor.

Technical Service

Providing excellent technical support for our products is one of our top priorities. Our technical support team relies on years of laboratory experi-ence to assist you with product selection or to help troubleshoot any issues that you may have. To contact our technical service representatives call either 1.888.208.2246 (toll free) or 1.512.707.8993, or email Bioo Scientific at [email protected].

Select Institutions Using NEXTflex™ Sequencing Kits & Reagents

Brown UnivCDCChildren’s Hospital BostonCold Spring Harbor LabColumbia Medical CenterDartmouth Medical SchoolFred Hutchison Cancer Research InstHarvard Medical SchoolJ. Craig Venter InstituteJohns Hopkins UnivMD Anderson Medical CenterMedical Neurogenetics Memorial Sloan Kettering Cancer CenterMIT

Mount Sinai School of MedicineNational Cancer InstituteNational Institute of HealthNew York UnivNorthern Arizona UnivOklahoma Medical Research FoundationRockefeller UnivSt Jude Children’s Research HospitalStowers InstituteTranslational Genomics Research Institute (TGEN)Univ of British ColumbiaUniv of California – BerkeleyUniv of California – DavisUniv of California – Riverside

Univ of California - San DiegoUniv of California – RiversideUniv of IllinoisUniv of MassachusettsUniv of Massachusetts Med SchoolUniv of MontanaUniv of N. CarolinaUniversity of Texas Health Science CenterUniv of WashingtonUniversity of Wisconsin, MadisonUSDAVanderbilt University

1.888.208.22461.512.707.8993

Our customer service representatives are ready to assist you with ordering Monday

through Friday

1.512.707.2051

Please include the following information with your order:

Catalog #, Name, Size & QuantityShipping/Billing Address

Name, Phone & Customer #Purchase Order # or Credit Info

[email protected]

Please include the following information with your order:

Catalog #, Name, Size & QuantityShipping/Billing Address

Name, Phone & Customer #Purchase Order # or Credit Info


IndexIndex

16S V4 Amplicon-Seq Kit

AIR™ DNA-Seq Barcodes - Illumina®

Bisulfite-Seq Barcodes - Illumina®

Bisulfite-Seq Kits - Illumina®

ChIP-Seq Kits - Illumina®

ChIP-Seq Barcodes - Illumina®

ChIP-Seq Modules - Illumina®

Directional RNA-Seq BarcodesDirectional RNA-Seq KitsDNA Fragmentation KitDNA Barcode Blockers - for SeqCap EZDNA Isolation KitDNA-Seq Barcodes - Illumina®

DNA-Seq Barcodes - Ion Proton™ & PGM™DNA-Seq Barcodes - 5500 SOLiD™DNA-Seq Barcodes - SOLiD™ 4DNA-Seq Kits - Illumina®

DNA-Seq Kit for Biomek - Illumina®

DNA-Seq Kits - Ion Proton™ & PGM™DNA-Seq Kits - 5500 SOLiD™DNA-Seq Kits - SOLiD™ 4DNA-Seq Modules - Illumina®

22

27

2616

112624

514558205926303436082330343624

08

131316

2618254720

40

0660514247

54614851

Enhanced Adapter Ligation Technology

MeDIP-MeCAP Kits - Illumina®

Methyl-Seq Kits - Illumina®

Msp 1 Restriction Enzyme

PCR-Free DNA-Seq Barcodes - Illumina®

PCR-Free DNA-Seq Kits - Illumina®

PCR-Free DNA-Seq Modules - Illumina®

Poly(A) BeadsPre-Capture Combo Kit

qRNA-Seq Kit - Illumina®

Rapid DNA-Seq KitsRNA Isolation KitRNA-Seq BarcodesRNA-Seq Kit - Illumina®

RNA-Seq Modules - Illumina®

SEQBOT™ Liquid HandlerSmall RNA Isolation KitSmall RNA-Seq KitsSmall RNA-Seq Barcodes

Next Generation Sequencing Catalog2014-2015

®

New RapidDNA-Seq Kitsee page 06

New 16S V4Amplicon-Seq Kit

see page 22

NewqRNA-Seq Kitsee page 40

Documents

Next Generation Sequencing Catalog Scientific NGS Catalog 2014... · 2015-07-02 · Introduction 2 Bioo Scientific Bioo Scientific is a global life science company headquartered in