High-throughput purification products from MACHEREY … · MN guide to high-throughput ... Elution (low-salt): Low-salt or water elution ... • Extensive troubleshooting by MN experts

MACHEREY-NAGEL GmbH & Co. KG · Neumann-Neander-Str. 6–8 · 52355 Düren · GermanySwitzerland:MACHEREY-NAGEL AGTel.: +41 62 388 55 00Fax: +41 62 388 55 05E-mail: [email protected]

Germanyand international:Tel.: +49 24 21 969-0Fax: +49 24 21 969-199E-mail: [email protected]

France:MACHEREY-NAGEL EURLTel.: +33 388 68 22 68Fax: +33 388 51 76 88E-mail: [email protected]

www.mn-net.com

www.mn-net.comMACHEREY-NAGEL

EN ISO 9001: 2008CERTIFIED

USA:MACHEREY-NAGEL Inc.Tel.: +1 484 821 0984Fax: +1 484 821 1272E-mail: [email protected]

HTP

Pur

ifica

tion

Gui

deHigh-throughput purification products from MACHEREY-NAGEL

MN guide to high-throughput (HTP) purificationFlexible solutions for successNucleoSpin®

NucleoFast®

NucleoMag®

High reliability in quality and quantityVersatility of throughput and purification technologyHigh value automated and manual processingMN expertise with outstanding technical support

www.mn-net.com2

MACHEREY-NAGEL HTP Kit Finder

NucleoSpin® 8 Plasmid

NucleoSpin® 8 Plasmid Core Kit

NucleoSpin® 96 Plasmid

NucleoSpin® 96 Plasmid Core Kit

NucleoSpin® 96 Flash

NucleoSpin® 8 PCR Clean-up

NucleoSpin® 8 PCR Clean-up Core Kit

NucleoSpin® 96 PCR Clean-up

NucleoSpin® 96 PCR Clean-up Core Kit

NucleoFast® 96 PCR

NucleoMag® 96 PCR

NucleoSpin® 8 RNA

NucleoSpin® 8 RNA Core Kit

NucleoSpin® 96 RNA

NucleoSpin® 96 RNA Core Kit

NucleoMag® 96 RNA

NucleoSpin® 8 Virus

NucleoSpin® 8 Virus Core Kit


NucleoSpin® 96 Virus Core Kit

NucleoMag® 96 Virus

Silicamembrane

Bacteria,yeast

PCR reaction

Cells, tissue, bacteria,

yeast, plant

Cell-free body fluids

Plasmid

Clean-up

RNA

ViralRNA/DNA

Silicamembrane

Ultra-filtration

Magnetic beads*

Silicamembrane

Magneticbeads*

Silicamembrane

Magneticbeads*

8-well 96-well

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Man

ual,

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* Manual processing of NucleoMag® kits is possible. A suitable magnetic separator is required (e.g., NucleoMag® SEP).

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MACHEREY-NAGEL HTP Kit Finder

NucleoSpin® 8 Blood

NucleoSpin® 8 Blood Core Kit

NucleoSpin® 8 Blood QuickPure


NucleoSpin® 96 Blood Core Kit


NucleoMag® Blood 200 μL, 3mL

NucleoSpin® 8 Tissue

NucleoSpin® 8 Tissue Core Kit


NucleoSpin® 96 Tissue Core Kit

NucleoMag® 96 Tissue

NucleoSpin® 8 Trace


NucleoMag® 96 Trace

NucleoSpin® 8 Plant II

NucleoSpin® 8 Plant II Core Kit


NucleoSpin® 96 Plant II Core Kit

NucleoMag® 96 Plant

NucleoSpin® 8 Food

NucleoSpin® 96 Food

Silicamembrane

Blood

GenomicDNA

Tissue,cells

Forensicsamples

Plant,fungi

Food,feed

Magneticbeads*

Silicamembrane

Magnetic beads*

Silicamembrane

Magneticbeads*

Silicamembrane

Magneticbeads*

Silicamembrane

8-well 96-well

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Man

ual,

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trifu

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Man

ual,

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um

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omat

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* Manual processing of NucleoMag® kits is possible. A suitable magnetic separator is required (e.g., NucleoMag® SEP).

www.mn-net.com4

Technologies

NucleoSpin® technologyTechnology / Material Silica-membrane Format 8-well strips Medium-throughput systems, for vacuum manifolds,

centrifuges, and robotic systems96-well plates High-throughput systems, for vacuum manifolds, centri-

fuges, and robotic systemsPrinciple Chaotropic salt binding (high-salt): Interaction between DNA / RNA

and silica membrane (hydrate shell is reversibly removed by chaotropic salt)Washing: High-salt and ethanolic washing steps remove contaminantsElution (low-salt): Low-salt or water elution (hydrate shell is recovered, DNA / RNA is released from the membrane)

Features / Result • High-quality, sequencing-grade and PCR-grade DNA / RNA• From medium to high-throughput • No alcohol precipitation necessary• Easy to use

NucleoMag® technologyTechnology / Material Superparamagnetic beads (non-silica)Format FlexiblePrinciple Chaotropic salt binding (high-salt): Adsorption of DNA and RNA to

the beads (hydrate shell is reversibly removed by chaotropic salt)Washing: High-salt and ethanolic washing steps remove contaminantsElution (low-salt): Low-salt or water elution (hydrate shell is recovered, DNA / RNA is released from the beads)

Features / Result Easily adapted to automated use due to:• Efficient bead separation• Easy resuspension and mixing• Minimized bead carry-over• Flexible throughput• Flexible sample volume• Minimized risk of cross-contamination (one-tube procedure)

NucleoFast® technologyTechnology / Material Ultrafiltration membraneFormat 96-well plates

High-throughput systems, for vacuum manifolds, centrifuges, and robotic systems

Principle UltrafiltrationRetention of DNA fragments > 150 bpOptional washing step with water

Features / Result • Fast and easy-to-use procedure• Pure, ready-to-use DNA• Detergent-free membrane• Membrane resistant against tip-touching• Cost-efficient

DNA / RNA is bound to the silica membrane under high-salt conditions

Contaminants are washed away under high-salt conditions

DNA / RNA is eluted using low-salt buffer or water

NucleoMag® Beads are added to the sample

DNA / RNA is bound to the NucleoMag® BeadsBeads are restrained in the well by the magnet while contaminants are washed away

DNA / RNA is eluted from the beads and recovered, while beads are re-strained in the well by the magnet

Sample is loaded directly onto the NucleoFast® Filter

Sample is collected on the surface of the ultrafiltration membrane while contaminants are filtered to waste

Purified DNA is recovered from the membrane after addition of water or buffer and a short incubation

DNA / RNA Contaminants NucleoMag® Beads

DNA / RNAContaminants

DNA / RNAContaminants

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Equipment

NucleoSpin® kitsFormats • NucleoSpin® kits are available in unique 8-well strips for

medium-throughput and in convenient 96-well plates for high-throughput.

• The maximum volume capacity of NucleoSpin® 8-well strips and 96-well plates is 1.4 mL.

• Optimized column outlets minimize the risk of cross- contamination by reducing spray during filtration and elution steps.

Automation In addition to our standard medium- and high-throughput kits, MN offers NucleoSpin® Core Kits optimized to be used on au-tomation platforms. They contain binding plates, necessary buffers and enzymes. Additional accessories (e.g., plastics) can be combined and ordered separately as needed.

Centrifugation Required specifications for centrifugation: • Microtiterplate centrifuge with a bucket height of up to 85 mm*• Centrifugal forces of 5600–6000 x g are needed.• For processing 8-well strips Starter Set C is required

(containing two Column Holders C, MN Square-well Blocks, Racks of Tube Strips).

Vacuum processing

• The unique MN Wash Plate prevents the bottom of the NucleoSpin® Binding Strips / Plates from being contami-nated by ethanolic wash buffers. (MN Wash plates are not included in NucleoSpin® Core Kits).

• For processing 8-well strips Starter Set A is required (containing two Column Holders A, 12 NucleoSpin® Dummy Strips).

• Required specifications for suitable vacuum manifolds: > 100 mbar; airflow 1800 L/h

• Compatible vacuum manifolds: NucleoVac 96 Vacuum Ma-nifold (MN), QIAvac 96 (QIAGEN), Vac-Man® 96 (Promega)

NucleoFast® kitsCentrifugation and vacuum processing

NucleoFast® kits can be processed by centrifugation or vacu-um. For processing NucleoFast® kits by centrifugation a suit-able waste collection plate is required, e.g., MN Square-well Block, and a microtiterplate centrifuge with centrifugal forces of 4500 x g.

NucleoMag® kitsAutomated and manual processing

NucleoMag® kits are predominantly used on automation plat-forms. For manual processing a magnetic separator is requi-red, e.g., NucleoMag® SEP. NucleoMag® SEP can also be integrated on automation platforms. For further details ple-ase inquire with Technical Service. NucleoMag® kits prima-rily contain magnetic beads, enzymes and buffers. Plastics have to be supplied separately according to the requirements of the individual automation platform. For an assortment of suitable consumables please refer to HTP Consumables on page 18.

* Height depends on application. Please inquire with Technical Service.

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Support

Suitable robotic platformsFor an overview of the robotic platforms that are suitable with MACHEREY-NAGEL kits visit: www.mn-net.com/robot-overview

Application notesMN offers a broad range of application notes. These application notes contain detailed descriptions on how to use medium- and high-throughput kits from MN on different robotic platforms. The number of available application notes increases continuously.For detailed information please visit:www.mn-net.com/HTP-application-notes

Personal support by MACHEREY-NAGEL experts• MN supplies validated and released basic scripts on request.• Extensive troubleshooting by MN experts.• Specialists from R&D assist you to generate customized scripts for different robotic platforms.• MN experts help to optimize or adjust your scripts on request, e.g., to process new sample material.• In order to benefit from these special services please use the inquiry form:

www.mn-net.com/HTP-inquiry-form or contact our Technical Support and Customer Service or Product Management:

Technical Support and Customer Service Tel.: +49 24 21 969-271

+49 24 21 969-270E-mail: [email protected]

Product Management HTPTel.: +49 24 21 969-271E-mail: [email protected]

www.mn-net.com 7

JJNucleoSpin® 8 / 96 Plasmid · NucleoSpin® 8 / 96 Plasmid Core Kit Silica membrane-based plasmid DNA purification

XX Suitable for up to 5 mL bacterial cultureXX Convenient filtration of bacterial lysates

Optimized filter material minimizes the risk of filter clogging

Page: 2 / 326.03.2010

Sequence: 359B2_M13uni Clipped length:Left clip:Right clip:Avg. qual. in clip.:

99412100551.92

Samples:Bases:Average spacing:

16301124214.0

Average quality >= 10: 99, 20: 92, 30: 885

Clip. 1 BQ 20 WL 10 Quality: 0 - 910 - 1920 - 29>= 30

T A G C T T G A G T510

A T T C T A T A G T520

G T C A C C T A A A530

T A G C T T G G C G540

T A A T C A T G G T550

C A T A G C T G T T560

T C C T G T G T G A570

A A T T G T T A T C580

C G C

T C A C A A T590

T C C A C A C A A C600

A T A C G A G C C G610

G A A G C A T A A A620

G T G T A A A G C C630

T G G G G T G C C T640

A A T G A G T G A G650

C T A A C T C A C A660

T T A A T T G

C G T670

T G C G C T C A C T680

G C C C G C T T T C690

C A G T C G G G A A700

A C C T G T C G T G710

C C A G C T G C A T720

T A A T G A A T C G730

G C C A A C G C G C740

G G G G A G A G G C750

G G

T T T G C G T A760

T T G G G C G C T C770

T T C C G C T T C C780

T C G C T C A C T G790

A C T C G C T G C G800

C T C G G T C G T T810

C G G C T G C G G C820

G A G C G G T A T C830

A G C T C A C

T C A840

A A G G C G G T A A850

T A C G G T T A T C860

C A C A G A A T C A870

G G G G A T A A C G880

C A C G A A A G A A890

C A T G T G A G C A900

A A A G G C C A G C910

A A A A G G C C A G920

G A A C

C G T A A A930

A G G C C G C G T T940

G C T G G C G T T T950

T T C C A T A G G C960

T C C G C C C C C T970

G A C G A G C A T C980

A C A A A A A T C G990

A C G C T C A A G T1000

C A G A G G T

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Sequence: 359B2_M13uni Clipped length:Left clip:Right clip:Avg. qual. in clip.:

99412100551.92

Samples:Bases:Average spacing:

16301124214.0

Average quality >= 10: 99, 20: 92, 30: 885

Clip. 1 BQ 20 WL 10 Quality: 0 - 910 - 1920 - 29>= 30

G G G G G A T A T C10

T A T A G G G C G A20

T T G G G C C C G A30

C G T C G C A T G C40

T C C C G G C C G C50

C A T G G C G G C C60

G C G G G A A T T C70

G A T T T G G G G T80

G C

C T A A T G A G90

T G A G C T A A C T100

C A C A T T A A T T110

G C G T T G C G C T120

C A C T G C C C G C130

T T T C C A G T C G140

G G A A A C C T G T150

C G T G C C A G C T160

G C A T T A A

T G A170

A T C G G C C A A C180

G C G C G G G G A G190

A G G C G G T T T G200

C G T A T T G G G C210

G C T C T T C C G C220

T T C C T C G C T C230

A C T G A C T C G C240

T G C G C T C G G T250

C G T T C G G C T G260

C G G C G A G C G G270

T A T C A G C T C A280

C T C A A A G G C G290

G T A A T A C G G T300

T A T C C A C A G A310

A T C A G G G G A T320

A A C G C A G G A A330

A G A A

C A T G T G340

A G C A A A A G G C350

C A G C A A A A G G360

C C A G G A A C C G370

T A A A A A G G C C380

G C G T T G C T G G390

C G T T T T T C C A400

T A G G C T C C G C410

C C C C C T G

A C G420

A G C A T C A C A A430

A A A A T C A C T A440

G T G A A T T C G C450

G G C C G C C T G C460

A G G T C G A C C A470

T A T G G G A G A G480

C T C C C A A C G C490

G T T G G A T G C A500

Consistently high yields and high reading length in sequencingpGEM® was isolated from 1 mL E. coli DH 5α™ culture grown in LB-medium, using Eppendorf epMotion® 5075 and NucleoSpin® 96 Plasmid. DNA was quantified using absorbance measurement. DNA sequencing was performed using ABI PRISM® BigDye® chemistry and T7 primer. Sequence was determined using the ABI 3730 XL DNA analyzer. High-quality reading lengths of approximately 800 nt were obtained.

Product Preps Specification REF

NucleoSpin® 8 Plasmid 12 x 8 / 60 x 8 NucleoSpin® Plasmid Filter Strips, NucleoSpin® Plasmid Binding Strips, Culture Plates, MN Wash Plates, Racks of Tube Strips, buffers, RNase A

740621 / .5

NucleoSpin® 8 Plasmid Core Kit*

48 x 8 NucleoSpin® Plasmid Filter Strips, NucleoSpin® Plasmid Binding Strips, buffers, RNase A

740461.4

NucleoSpin® 96 Plasmid 1 x 96 / 4 x 96 / 24 x 96 NucleoSpin® Plasmid Filter Plates, NucleoSpin® Plasmid Binding Plates, MN Wash Plates, MN Square-well Blocks, Elution Plates U-bottom, Gas-permeable Foils, Self-ad-hering Foils, buffers, RNase A

740625.1 / .4 / .24

NucleoSpin® 96 Plasmid Core Kit*

4 x 96 NucleoSpin® Plasmid Filter Plates, NucleoSpin® Plasmid Binding Plates, buffers, RNase A

740616.4

* Core Kit contents are focused on automation platforms. Additional accessories can be combined as needed. Please check www.mn-net.com or contact our Technical Support ([email protected]).

1) Support protocol available. Please contact our Technical Support ([email protected]).2) Suitable centrifuge to harvest bacterial cells is required. For details see corresponding user manual at www.mn-net.com.

JJNucleoSpin® 96 Flash Fast high-throughput purification of plasmid DNA

XX Suitable for high-throughput purification of large constructs (cosmids, BACs)XX Vector size < 250 kbpXX Fast and cost-efficient procedureXX Suitable for 1–3.9 mL E. coli culture


NucleoSpin® 96 Flash 2 x 96 / 4 x 96 / 24 x 96 NucleoSpin® Flash Filter Plates, MN Square-well Blocks, Square-well Blocks, Gas-permeable Foils, Self-adhering Foils, buffers, RNase A

740618.2 / .4 / .24

1) Suitable centrifuge to harvest bacterial cells and to precipitate DNA is required. For details see corresponding user manual at www.mn-net.com.

Processing:• automated • manual, centrifuge1 • manual, vacuum2

MN Wash Plate !i

Processing:• automated1 • manual, centrifuge • manual, vacuum1 i

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JJNucleoSpin® 8 / 96 PCR Clean-up · NucleoSpin® 8 / 96 PCR Clean-up Core Kit Silica membrane-based PCR clean-up

XX Quantitative primer removal up to 100 ntXX High recovery for a wide range of fragment sizes (64 bp–10 kbp)

Recovery rates of even small fragments Yield of DNA fragments before (a) and DNA recovery after (b) purification with NucleoSpin® 96 PCR Clean-up. Samples of 25 μL were purified and analyzed on a 1 % TAE agarose gel.


NucleoSpin® 8 PCR Clean-up 12 x 8 / 60 x 8 NucleoSpin® PCR Clean-up Binding Strips, MN Wash Plates, Racks of Tube Strips, buffers

740668 / .5

NucleoSpin® 8 PCR Clean-up Core Kit*

48 x 8 NucleoSpin® PCR Clean-up Binding Strips, buffers 740463.4

NucleoSpin® 96 PCR Clean-up 1 x 96 / 2 x 96 / 4 x 96 / 24 x 96

NucleoSpin® PCR Clean-up Binding Plates, MN Wash Plates, Elution Plates U-bottom, Self-adhering Foils, buffers

740658.1 / .2 / .4 / .24

NucleoSpin® 96 PCR Clean-up Core Kit*

4 x 96 NucleoSpin® PCR Clean-up Binding Plates, buffers 740464.4

* Core kit contents are focused on automation platforms. Additional accessories can be combined as needed. Please check www.mn-net.com or contact our Technical Support ([email protected]).

1) Support protocol available. Please contact our Technical Support ([email protected]).

JJNucleoMag® 96 PCR Magnetic bead-based PCR clean-up

XX Highly concentrated DNA Small elution volumes down to 25 μL Yield is not affected by different elution volumes

XX Flexible sample volume

High recovery of even small PCR fragmentsPurification of different fragment sizes using NucleoMag® 96 PCR. Recovery depends on fragment length. On average, a recovery of 80–95 % is obtained.


NucleoMag® 96 PCR 1 x 96 / 4 x 96 / 24 x 96 NucleoMag® P-Beads, Elution Plates U-bottom, Self-adhering PE Foils, buffers

744100.1 / .4 / .24

Rec

over

y [%

]

100

60

20

bp 164 252 359 459 645 787 982 1484Recovery [%] 78 81 80 95 92 94 83 81

a b- 64 bp

- 164 bp

- 252 bp

- 359 bp

- 459 bp

- 645 bp

- 782 bp

- 982 bp

a b

Processing:• automated • manual, centrifuge1 • manual, vacuumMN Wash Plate !

i

Processing:• automated • manual i

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JJNucleoFast® 96 PCR Ultrafiltration-based PCR clean-up

XX Optimal membrane for ultrafiltration Detergent-free membrane, resistant against tip touching easy recovery of pure DNA

XX Suitable to concentrate pre-purified DNA / RNA > 150 bpXX Fast procedure and easy handlingXX Complete kits and separate plates available


NucleoFast® 96 PCR Clean-up Kit

4 x 96 NucleoFast® 96 PCR Plates, Elution Plates U-bottom, Self-adhering Foils, buffers

743500.4

NucleoFast® 96 PCR Plates

10 x 96 / 50 x 96 NucleoFast® 96 PCR Plates 743100.10 / .50

JJNucleoSpin® 8 / 96 RNA · NucleoSpin® 8 / 96 RNA Core Kit Silica membrane-based isolation of total RNA from tissue and cell samples

XX Highest purity Recombinant DNase for efficient on-column removal of genomic DNA RNA ready to use for all common downstream applications

XX No organic extraction steps

Total RNA of high structural integrityElectropherograms obtained from Agilent Bio-analyzer 2100 using RNA Nano 6000 reagents. RNA was isolated from cells (A) or frozen liver tissue (B). RNA integrity numbers (RIN) are 9.7 for RNA isolated from cells and 9.1 for RNA iso-lated from frozen liver tissue.


NucleoSpin® 8 RNA 12 x 8 / 60 x 8 NucleoSpin® RNA Binding Strips, MN Wash Plates, MN Square-well Blocks, Racks of Tube Strips, Elution Plates U-bottom, Self- adhering Foils, buffers, RNase-free rDNase

740698 / .5

NucleoSpin® 8 RNA Core Kit*

48 x 8 NucleoSpin® RNA Binding Strips, buffers, RNase-free rDNase 740465.4

NucleoSpin® 96 RNA 2 x 96 / 4 x 96 / 24 x 96 NucleoSpin® RNA Binding Plates, MN Wash Plates, MN Square-well Blocks, Round-well Blocks Low, Elution Plates U-bottom, Self-adhering Foils, buffers, RNase-free rDNase

740709.2 / .4 / .24

NucleoSpin® 96 RNA Core Kit*

4 x 96 NucleoSpin® RNA Binding Plates, buffers, RNase-free rDNase 740466.4

For sample homogenization and removal of cell debris it is recommended to filter the lysate through NucleoSpin® RNA Filter Strips (REF 740699.12F / 60F) or NucleoSpin® RNA Filter Plates (REF 740711). Filter strips or plates are not included in the kits.* Core Kit contents are focused on automation platforms. Additional accessories can be combined as needed.1) Suitable centrifuge to harvest cells grown in suspension is required. For details see corresponding user manual at www.mn-net.com.

A) RNA from cultured cells RIN: 9.7

B) RNA from frozen liver tissue RIN: 9.1

Processing:• automated • manual, centrifuge • manual, vacuum i

Processing:• automated • manual, centrifuge • manual, vacuum1

MN Wash Plate !i

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JJNucleoMag® 96 RNA Magnetic bead-based isolation of RNA from tissue and cell samples

XX Outstanding RNA purity and qualityXX Highly concentrated RNA

Small elution volumes ≥ 50 μL

XX Convenient sample processing No organic extraction steps Reducing agent TCEP included (β-mercaptoethanol not needed)


NucleoMag® 96 RNA 1 x 96 / 4 x 96 NucleoMag® B-Beads, buffers, TCEP, RNase-free rDNase 744350.1/.4

JJNucleoSpin® 8 / 96 Blood · NucleoSpin® 8 / 96 Blood Core Kit Silica membrane-based isolation of genomic DNA from body fluids

XX Convenient handling Improved flow-rate reduces risk of clogging Complete processing at room temperature

XX Reliable results: consistently high yields and purities

Reproducible yield and purity96 aliquots of 200 μL blood from the same sample have been purified fully automated on a Tecan Genesis™. The average yield is 6.76 ± 0.71 μg with a coefficient of variation (CV) of 10 %. The average purity is 1.97 ± 0.02 (CV: 1 %).


NucleoSpin® 8 Blood 12 x 8 / 60 x 8 NucleoSpin® Blood Binding Strips, MN Wash Plates, Racks of Tube Strips, buffers, Proteinase K

740664 / .5

NucleoSpin® 8 Blood Core Kit*

48 x 8 NucleoSpin® Blood Binding Strips, buffers, Proteinase K 740455.4


1 x 96 / 4 x 96 / 24 x 96

NucleoSpin® Blood Binding Plates, MN Wash Plates, Lysis Blocks, Racks of Tube Strips, Tubes (15 mL), buffers, Proteinase K

740665.1 / .4 / .24

NucleoSpin® 96 Blood Core Kit*

4 x 96 NucleoSpin® Blood Binding Plates, buffers, Proteinase K 740456.4


Processing:• automated • manual, centrifuge • manual, vacuumMN Wash Plate !

i


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JJNucleoSpin® 8 / 96 Blood QuickPure Silica membrane-based isolation of genomic DNA from body fluidsXX Time-saving parallel isolation of genomic DNA

Reduced number of washing and drying steps minimizes hands-on-time

XX Suitable for difficult blood samples (e.g., animal blood)XX Consistently high DNA yields

Reproducible yield and purityDNA was isolated with NucleoSpin® 96 Blood QuickPure from blood aliquots of a single blood donation (EDTA blood, 200 μL each) according to the proto-col. DNA yield was determined by absorbance measurement (Lambda Scan 200, MWG-Biotech). The average yield is 4.79±0.50 μg with a coefficient of variation (CV) of 10 %. The average purity is 1.94±0.08 (CV: 4 %).



12 x 8 / 60 x 8 NucleoSpin® Blood QuickPure Binding Strips, Racks of Tube Strips, Self-adhering Foils, buffers, Proteinase K

740666 / .5


2 x 96 / 4 x 96 / 24 x 96 NucleoSpin® Blood QuickPure Binding Plates, MN Square-well Blocks, Round-well Blocks Low, Self-adhering Foils, buffers, Proteinase K

740667.2 / .4 / .24

JJNucleoMag® Blood 200 μL • NucleoMag® Blood 3 mL Magnetic bead-based isolation of genomic DNA from body fluids

XX Scalable DNA isolation and convenient handling Two different kits to meet the need for different DNA yields Complete processing possible at room temperature

XX High concentration of high quality DNA Small elution volumes: ≥ 50 μL (NucleoMag® Blood 200 μL), ≥ 1 mL (NucleoMag® Blood 3 mL)

High consistency of DNA yield from fresh and frozen bloodSamples were taken from six different donors (S1-S6). Sample aliquots from each donor were purified either from fresh samples or after one day storage at -20 °C. The extraction was carried out 8 times per sample. The value given is the median of all 8 extractions. Highly reproducible yields were obtained from fresh or frozen blood samples.

Scalable DNA isolation from up to 3 mL human blood sampleIsolation of DNA from 100 μL to 3 mL blood using NucleoMag® Blood 3 mL and KingFisher® Flex 24. DNA yield is increased proportionally to the used sample volume.


NucleoMag® Blood 200 μL

1 x 96 / 4 x 96 NucleoMag® B-Beads, buffers, Proteinase K 744501.1 / .4

NucleoMag® Blood 3 mL

1 x 96 NucleoMag® B-Beads, buffers, Proteinase K 744502.1


Processing:• manual, centrifuge i

Yiel

d [μ

g]

Yiel

d [μ

g]

Sample volume [mL]

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JJNucleoSpin® 8 / 96 Tissue · NucleoSpin® 8 / 96 Tissue Core Kit Silica membrane-based isolation of genomic DNA from human or animal tissue, cultured cells,

or bacteria

XX Highly pure DNA Efficient removal of PCR inhibitors Ready to use for genotyping analysis

XX Reliable DNA extraction for consistent downstream analysis

Real time PCR of DNA isolated from mouse tailsDNA isolated from mouse tails was used in a SYBR® Green PCR assay. PCR amplification plots show reproducible amplification. The average crossing point for all sixteen samples was 23.79 with a CV of 2.43 %.

DNA isolation with NucleoSpin® 8 Tissue from different kinds of mouse tissue samplesDNA was isolated from approximately 20 mg of the indicated tissues. Each bar represents the average of eight extractions. Error bars indicate the standard deviation for each set of extractions.



12 x 8 / 60 x 8 NucleoSpin® Tissue Binding Strips, MN Wash Plates, MN Square-well Blocks, Rack of Tube Strips, Self-adhering Foils, buffers, Proteinase K

740740 / .5

NucleoSpin® 8 Tissue Core Kit*

48 x 8 NucleoSpin® Tissue Binding Strips, buffers, Proteinase K 740453.4


2 x 96 / 4 x 96 / 24 x 96

NucleoSpin® Tissue Binding Plates, MN Wash Plates, MN Square-well Blocks, Round-well Blocks, Racks of Tube Strips, Self-adhering Foils, buffers, Proteinase K

740741.2 / .4 / .24

NucleoSpin® 96 Tissue Core Kit*

4 x 96 NucleoSpin® Tissue Binding Plates, buffers, Proteinase K 740454.4


1) Suitable centrifuge to clear, e.g., lysates from mouse or rat tails is required. For details see corresponding user manual at www.mn-net.com.

JJNucleoMag® 96 Tissue Magnetic bead-based isolation of genomic DNA from human or animal tissue, cultured cells,

or bacteria

XX Reliable DNA purification for reproducible results Highly pure DNA ready to use for genotyping applications

XX Consistent DNA yield Small elution volumes ≥ 50 μL possible Yield is not affected by different elution volumes

XX Processing at room temperature (excl. lysis step)


NucleoMag® 96 Tissue

1 x 96 / 4 x 96 / 24 x 96

NucleoMag® B-Beads, buffers, Proteinase K 744300.1 / .4 / .24


MN Wash Plate !i


www.mn-net.com 13

JJNucleoSpin® 8 / 96 Trace Silica membrane-based isolation of genomic DNA from forensic samples

XX Isolation of genomic DNA from various forensic samples (e.g., buccal swabs, dried blood spots, cigarette butts)XX DNA ready to use for, e.g., PCR, STR analysis, any kind of enzymatic reaction

STR analysisGenomic DNA has been prepared from buccal swabs using NucleoSpin® 8 Trace. DNA was quantified with Quantifiler® Human DNA Quantification Kit (Applied Biosystems) on 7500 Thermocycler (applied biosystems). 750 pg DNA has been used in a Multiplex PCR using the AmpFlSTR® NGM™ PCR Amplification Kit (Applied Biosystems). Se-quencing and subsequent analysis was performed on a 3500 Genetic Analyzer with GeneMapper® ID-X 1.0 software (Applied Biosystems).(Data kindly provided by Dr. Witt, Landeskriminalamt Hamburg, Germany.)



12 x 8 / 60 x 8 NucleoSpin® Trace Binding Strips, MN Wash Plates, MN Square-well Blocks, Racks of Tube Strips, Self-adhering Foils, buffers, Proteinase K

740722.1 / .5


2 x 96 / 4 x 96 NucleoSpin® Trace Binding Plates, MN Wash Plates, MN Square-well Blocks, Racks of Tube Strips, buffers, Proteinase K

740726.2 / .4

To separate the DNA containing lysate from the forensic material the NucleoSpin® Trace Filter Plate (REF 740677) is recommended. NucleoSpin® Trace Filter Plates are not included in the kit.1) Suitable centrifuge for separation of lysate and sample material and for elution is recommended. For details see corresponding user manual at www.mn-net.com.

JJNucleoMag® 96 Trace Magnetic bead-based isolation of genomic DNA from forensic samples

XX Isolation of genomic DNA from various forensic samples (e.g., buccal swabs, dried blood spots, cigarette butts) XX DNA ready to use for, e.g., PCR, STR analysis, any kind of enzymatic reactionXX Highly concentrated DNA

Small elution volumes ≥ 50 μl possible Yield is not affected by different elution volumes


NucleoMag® 96 Trace

1 x 96 / 4 x 96 / 24 x 96

NucleoMag® B-Beads, buffers, Proteinase K 744600.1 / .4 / .24


MN Wash Plate !i


www.mn-net.com14

JJNucleoSpin® 8 / 96 Plant II · NucleoSpin® 8 / 96 Plant II Core Kit Silica membrane-based isolation of genomic DNA from plant cells and tissue

XX Broad range of starting materials can be processed Two different lysis buffers included: PL1 (CTAB lysis method) and PL2 (SDS lysis method)

XX Optimized DNA yield and purity New special silica membrane New wash buffer chemistry

XX RNase A included

DNA isolation from wheat leaves using NucleoSpin® 96 Plant IIReproducible isolation of genomic DNA from a master lysate represent-ing 50 mg of frozen wheat leaf tissue per extraction (n=48, identical sample lysate for all extractions) according to the standard procedure of NucleoSpin® 96 Plant II. The average yield is 10.5±0.5 μg with a coef-ficient of variation (CV) of 4.8 %.



12 x 8 / 60 x 8 NucleoSpin® Plant II Binding Strips, MN Wash Plates, MN Square-well Blocks, Racks of Tube Strips, Gas-permeable Foils, Self-adhering Foils, buffers, RNase A

740669 / .5

NucleoSpin® 8 Plant II Core Kit*

48 x 8 NucleoSpin® Plant II Binding Strips, buffers, RNase A 740467.4


2 x 96 / 4 x 96 / 24 x 96

NucleoSpin® Plant II Binding Plates, MN Wash Plates, MN Square-well Blocks, Racks of Tube Strips, Gas-permeable Foils, buffers, RNase A

740663.2 / .4 / .24

NucleoSpin® 96 Plant II Core Kit*

4 x 96 NucleoSpin® Plant II Binding Plates, buffers, RNase A 740468.4


1) Suitable centrifuge for lysate clearing is required. For details see corresponding user manual at www.mn-net.com.

JJNucleoMag® 96 Plant Magnetic bead-based isolation of genomic DNA from plant tissue

XX Easily scalable in sample volumeXX Highly concentrated DNA

Small elution volumes ≥ 50 μL possible Yield is not affected by different elution volumes


NucleoMag® 96 Plant

1 x 96 / 4 x 96 / 24 x 96

NucleoMag® C-Beads, Elution Plates U-bottom, buffers, RNase A 744400.1 / .4 / .24


MN Wash Plate !i


www.mn-net.com 15

JJNucleoSpin® 8 / 96 Virus · NucleoSpin® 8 / 96 Virus Core Kit Silica membrane-based isolation of viral RNA / DNA from serum, plasma, or cell-free biological

fluids

XX Quantitative extraction of viral RNA / DNA for a broad range of virus titersXX Simultaneous isolation of RNA and DNA

Proteinase K and Carrier RNA included

Highly sensitive detection of viral RNAHCV RNA was purified from 96 plasma samples (100 μL). A dilution series was processed using NucleoSpin® 96 Virus (12 samples each). Percentage of positive RT-PCR samples at low virus titers are shown (COBAS Amplicor, Roche Diagnostics).



12 x 8 / 60 x 8 NucleoSpin® Virus Binding Strips, MN Square-well Blocks, Racks of Tube Strips, Self-adhering Foils, buffers, Carrier RNA, Proteinase K

740643 / .5

NucleoSpin® 8 Virus Core Kit*

48 x 8 NucleoSpin® Virus Binding Strips, buffers, Carrier RNA 740451.4


2 x 96 / 4 x 96 NucleoSpin® Virus Binding Plates, MN Square-well Blocks, Round-well Blocks, Racks of Tube Strips, Self-adhering Foils, buffers, Carrier RNA, Proteinase K

740691.2 / .4

NucleoSpin® 96 Virus Core Kit*

4 x 96 NucleoSpin® Virus Binding Plates, buffers, Carrier RNA 740452.4


JJNucleoMag® Virus Magnetic bead-based isolation of viral RNA and DNA from serum or plasma

XX Quantitative extraction of viral RNA / DNA for a broad range of virus titersXX Simultaneous isolation of RNA and DNA

Proteinase K and Carrier RNA included

XX Highly concentrated RNA and DNA Small elution volumes ≥ 50 μL possible Yield is not affected by different elution volumes

H1N1 RNA detection from swab samplesIsolated viral RNA from swab samples (using NucleoMag® 96 Virus) was subjected to RT-RCR reactions specific for H1N1.RT-PCR setup was performed according to the recommendations of the Robert-Koch Institute (RKI) on a Roche LightCycler® 480 instrument. All results are in compliance with the reference controls.


NucleoMag® 96 Virus

1 x 96 / 4 x 96 NucleoMag® V-Beads, buffers, Carrier RNA, Proteinase K 744800.1 / .4

Fluo

resc

ence

(465

– 5

10)

Cycles

28.09725.59723.09720.59718.09715.59713.09710.597

8.0975.5973.0970.597

5 10 15 20 25 30 35 40 45 50

Processing:• automated • manual, centrifuge • manual, vacuumMN Wash Plate !

i


www.mn-net.com16

JJNucleoSpin® 8 / 96 Food Silica membrane-based isolation of genomic DNA from food and feed

XX Suitable for sensitive downstream applications, such as real-time PCR Complete removal of PCR inhibitors

XX Successful lysis of complex matrices, e.g., maize, cereals, biscuits, meat etc. Proprietary lysis buffer CF for food matrices

Isolation of genomic DNA from different kinds of shredded grainsGenomic DNA was isolated from 200 mg shredded wheat, sunflower seeds, grains of maize and soy. Indicated aliquots have been taken from 120 μL eluates and analyzed on a 1 % agarose gel (marker M: Lambda HindIII). The average yield for wheat, sunflower, maize and soy was 10.28 μg, 4.77 mg, 0.58 μg and 2.54 μg.


NucleoSpin® 8 Food

12 x 8 / 60 x 8 NucleoSpin® Food Binding Strips, MN Wash Plates, Round-well Blocks, Racks of Tube Strips, Gas-permeable Foils, buffers, Proteinase K

740975 / .5

NucleoSpin® 96 Food

2 x 96 / 4 x 96 / 24 x 96 NucleoSpin® Food Binding Plates, MN Wash Plates, MN Square-well Blocks, Round-well Blocks, Racks of Tube Strips, Gas-permeable Foils, buffers, Proteinase K

740976.2 / .4 / .24

1) Suitable centrifuge for lysate clearing is required. For details see corresponding user manual at www.mn-net.com.


MN Wash Plate !i

M Wheat M

M Sunflower M

M Maize M

M Soy M

10 μL Eluate

20 μL Eluate

20 μL Eluate

30 μL Eluate

www.mn-net.com 17

HTP equipmentProduct Pack of Specification REF

NucleoVac 96 Vacuum Manifold

1 Vacuum manifold, consists of manifold base and lid, a spacer set, and a waste container set Starter Set A is required when using NucleoSpin® 8-well strips on NucleoVac 96 (see below)

740681

NucleoVac Vacuum Regulator

1 For controlling of vacuum 740641

NucleoSpin® Dummy Strips

6 strips For closing unused rows of Column Holders A and B during vacuum processing of NucleoSpin® 8-well kits

740685

MN Frame 1 For optimized handling of 96-well plates with vacuum manifold on BioRobot® 9600, 9604, and 3000 (Qiagen), MultiPROBE® II / Janus (PerkinElmer), Biomek® 2000 / 3000 and FX / NX (Beckman Coulter)

740680

NucleoMag® SEP 1 Magnetic separator, for use with 96-well plates (e.g., REF 740481)

744900

Rubber Pad 2 Reusable mat to cover unused rows of a 96-well plate 740640

Column Holder A

Starter Set A 1 For processing NucleoSpin® 8-well strips under vacuum on NucleoVac 96 Vacuum Manifold or similar manifoldsContains 2 Column Holders A, NucleoSpin® Dummy Strips

740682

Column Holder B

Column Holder D

Starter Set B 1 For processing NucleoSpin® 8-well strips on the Qiagen BioRobot® 9600 / 9604 / 3000 Contains 1 Column Holder B, 1 Column Holder D,NucleoSpin® Dummy Strips

740683

Column Holder C

Starter Set C 1 For processing NucleoSpin® 8-well strips under centrifugationContains 2 Column Holders C, MN Square-well Blocks,Racks of Tube Strips

740684

www.mn-net.com18

HTP consumablesProduct Pack of Specification REF

MN Wash Plate 4

24

To facilitate drying of the DNA / RNA binding module and to minimize the risk of cross-contaminationAs above

740479

740479.24Square-well Block 4

24For use with NucleoMag® SEP (744900)As above

740481740481.24

MN Square-well Block 424

96-well blocks with 2.1 mL square wellsAs above

740476740476.24

Culture Plate 4 sets

24 sets

Square-well Block with 2.1 mL square wells used for growing, harvesting, or lysing of bacterial cultures, with Gas-permeable FoilAs above

740488 740488.24

Round-well Block

Round-well Block with Cap Strips

20

4 sets 24 sets

96-well blocks with 1.2 mL round wells used for, e.g., mixing steps and collecting elution fractions Set consists of 1 Round-well Block and 12 Cap Strips As above

740671

740475740475.24

Round-well Block Low 44 sets 24 sets

96-well blocks with 0.8 mL round wells96-well blocks with 0.8 mL round wells and Self-adhering FoilAs above

740482740487 740487.24

Elution Plate U-bottom 24 96-well microplates with 300 μL u-bottom wells with Self-adhering Foil

740486.24

Rack of Tube Strips 5 sets4 sets24 sets

1 set consists of 1 rack, 12 strips with 8 tubes each1 set consists of 1 rack, 12 strips with 8 tubes each, 12 cap stripsAs above

740637740477 740477.24

Cap Strips 48288

For sealing of Tube Strips, Round-well Blocks As above

740478740478.24

Gas-permeable Foil 50 To cover square-well blocks during incubation of bacterial cultures 740675

Self-adhering PE Foil 50 Adhesive tape foils for sealing 96-well elution plates for airtight storage of DNA / RNA

740676

NucleoSpin® Plasmid Filter Strips 48 For clarification of lysates, for use under vacuum or centrifugation 740730.48F

NucleoSpin® RNA Filter Strips 12

60

For filtration of cell and tissue homogenates, for use under vacuum or centrifugationAs above

740699.12F

740699.60FNucleoSpin® RNA Filter Plate 4 For filtration of cell and tissue homogenates, for use under vacuum

or centrifugation740711

NucleoSpin® Trace Filter Plate 20 For lysis of samples and subsequent removal of particulate matter, for use under vacuum or centrifugation

740677

Receiver Plates 20 μm Receiver Plates 20 μm hydrophilized

4

4

96-well plates with inserted filter frits of 20 μm pore size, suitable for centrifugation and vacuum 96-well plates with inserted hydrophilized filter frits of 20 μm pore size, suitable for gravity flow, centrifugation, and vacuum

740686.4

740687.4

Receiver Plates 50 μmReceiver Plates 50 μm hydrophilized

4

4

96-well plates with inserted filter frits of 50 μm pore size, suitable for centrifugation and vacuum 96-well plates with inserted hydrophilized filter frits of 50 μm pore size, suitable for gravity flow, centrifugation, and vacuum

740688.4

740689.4

KingFisher® 96 Accessory Kit A 1 set Square-well Blocks, Deep-well Tip Combs, Elution Plates, for 4 x 96 NucleoMag® 96 Tissue / Trace / Virus preps using KingFisher® 96 platform

744950

KingFisher® 96 Accessory Kit B 1 set Square-well Blocks, Deep-well Tip Combs, Elution Plates, for 4 x 96 NucleoMag® Blood 200 μL and NucleoMag® 96 Plant / RNA preps using KingFisher® 96 platform

744951

www.mn-net.com 19

Ordering informationProduct* Preps REFPlasmid DNA

NucleoSpin® 8 PlasmidNucleoSpin® 8 Plasmid Core KitNucleoSpin® 96 PlasmidNucleoSpin® 96 Plasmid Core Kit

12 x 8 / 60 x 848 x 81 x 96 / 4 x 96/24 x 964 x 96

740621 / .5740461.4740625.1 / .4 / .24740616.4

NucleoSpin® 96 Flash 2 x 96 / 4 x 96 / 24 x 96 740618.2 / .4 / .24PCR clean-up

NucleoSpin® 8 PCR Clean-upNucleoSpin® 8 PCR Clean-up Core KitNucleoSpin® 96 PCR Clean-upNucleoSpin® 96 PCR Clean-up Core Kit

12 x 8 / 60 x 848 x 81 x 96 / 2 x 96 / 4 x 96 / 24 x 964 x 96

740668 / .5740463.4740658.1 / .2 / .4 / .24740464.4

NucleoFast® 96 PCR Clean-up KitNucleoFast® 96 PCR Plates

4 x 9610 x 96 / 50 x 96

743500.4743100.10 / .50

NucleoMag® 96 PCR 1 x 96 / 4 x 96 / 24 x 96 744100.1 / .4 / .24Total RNA from cells and tissue

NucleoSpin® 8 RNANucleoSpin® 8 RNA Core KitNucleoSpin® 96 RNANucleoSpin® 96 RNA Core Kit

12 x 8 / 60 x 848 x 82 x 96 / 4 x 96 / 24 x 964 x 96

740698 / .5740465.4740709.2 / .4 / .24740466.4

NucleoMag® 96 RNA 1 x 96 / 4 x 96 744350.1 / .4Genomic DNA from blood

NucleoSpin® 8 BloodNucleoSpin® 8 Blood Core KitNucleoSpin® 96 BloodNucleoSpin® 96 Blood Core Kit

12 x 8 / 60 x 848 x 81 x 96 / 4 x 96 / 24 x 964 x 96

740664 / .5740455.4740665.1 / .4 / .24740456.4

NucleoSpin® 8 Blood QuickPureNucleoSpin® 96 Blood QuickPure

12 x 8 / 60 x 82 x 96 / 4 x 96 / 24 x 96

740666 / .5740667.2 / .4 / .24

NucleoMag® Blood 200 μLNucleoMag® Blood 3 mL

1 x 96 / 4 x 961 x 96

744501.1 / .4744502.1

Genomic DNA from tissue and cells

NucleoSpin® 8 TissueNucleoSpin® 8 Tissue Core KitNucleoSpin® 96 TissueNucleoSpin® 96 Tissue Core Kit

12 x 8 / 60 x 848 x 82 x 96 / 4 x 96 / 24 x 964 x 96

740740 / .5740453.4740741.2 / .4 / .24740454.4

NucleoMag® 96 Tissue 1 x 96 / 4 x 96 / 24 x 96 744300.1 / .4 / .24Genomic DNA from forensic samples

NucleoSpin® 8 TraceNucleoSpin® 96 Trace

12 x 8 / 60 x 82 x 96 / 4 x 96

740722.1 / .5740726.2 / .4

NucleoMag® 96 Trace 1 x 96 / 4 x 96 / 24 x 96 744600.1 / .4 / .24Genomic DNA from plant and fungi

NucleoSpin® 8 Plant IINucleoSpin® 8 Plant II Core KitNucleoSpin® 96 Plant IINucleoSpin® 96 Plant II Core Kit

12 x 8 / 60 x 848 x 82 x 96 / 4 x 96 / 24 x 964 x 96

740669 / .5740467.4740663.2 / .4 / .24740468.4

NucleoMag® 96 Plant II 1 x 96 / 4 x 96 / 24 x 96 744400.1 / .4 / .24Genomic DNA from food and feed

NucleoSpin® 8 FoodNucleoSpin® 96 Food

12 x 8 / 60 x 82 x 96 / 4 x 96 / 24 x 96

740975 / .5740976.2 / .4 / .24

Viral RNA and DNA from serum and plasma

NucleoSpin® 8 VirusNucleoSpin® 8 Virus Core KitNucleoSpin® 96 VirusNucleoSpin® 96 Virus Core Kit

12 x 8 / 60 x 848 x 82 x 96 / 4 x 964 x 96

740643 / .5740451.4740691.2 / .4740452.4

NucleoMag® 96 Virus 1 x 96 / 4 x 96 744800.1 / .4 *Kits to be used for research purposes only.

MACHEREY-NAGEL GmbH & Co. KG · Neumann-Neander-Str. 6–8 · 52355 Düren · GermanySwitzerland:MACHEREY-NAGEL AGTel.: +41 62 388 55 00Fax: +41 62 388 55 05E-mail: [email protected]

Germanyand international:Tel.: +49 24 21 969-0Fax: +49 24 21 969-199E-mail: [email protected]

France:MACHEREY-NAGEL EURLTel.: +33 388 68 22 68Fax: +33 388 51 76 88E-mail: [email protected]

www.mn-net.com

www.mn-net.comMACHEREY-NAGEL

EN ISO 9001: 2008CERTIFIED

USA:MACHEREY-NAGEL Inc.Tel.: +1 484 821 0984Fax: +1 484 821 1272E-mail: [email protected]

Your local distributor

Service Bioanalysis

Technical Support and Customer Service Tel.: +49 24 21 969-270 E-mail: [email protected] Management HTP Tel.: +49 24 21 969-271 E-mail [email protected]

Visit our Bioanalysis Pages at www.mn-net.com

Trademarks MACHEREY-NAGEL: NucleoSpin®, NucleoFast®, NucleoMag®

Other companiesApplied Biosystems: PRISM®, Quantifiler®, AmpFlSTR®, NGM™, GeneMapper®

Eppendorf: epMotion®

Tecan: Genesis Freedom®

Thermo Fisher Scientific Inc.: KingFisher®

Molecular Probes Inc.: SYBR®

Applera Corp.: Big Dye™Roche: LightCycler® Qiagen GmbH: BioRobot® Perkin Elmer LAS Inc.: MultiPROBE®

Beckman Coulter Inc.: Biomek®

Promega: Vac-Man®, pGEM®

Invitrogen: DH5α™

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