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Improving the Performance of SureSelectXT2 Target Capture
What is SureSelectXT2 Target Capture?
A solution-based hybridization target capture system that enriches
for regions of interest, as opposed to whole-genome sequencing
A wide range of predesigned probesets or baits and flexible customization
options allows users to design theirown capture panels
To see how it works, next we’ll take a look at the target capture protocol
0101 DNA ISOLATION
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DNA ISOLATION
LIBRARY PREP WITH BARCODES
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DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
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DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
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DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
WASH & RECOVER CAPTURED DNA
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DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
AMPLIFY CAPTURED DNA
WASH & RECOVER CAPTURED DNA
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DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
AMPLIFY CAPTURED DNA
WASH & RECOVER CAPTURED DNA
QUANTITATE USING qPCR
Co
ntain
ed
in B
ioo
Scie
ntifi
c’s Kit
Now that we’ve seen the protocol, let’s focus on Step 3: HYBRIDIZE WITH BLOCKERS
0303 HYBRIDIZE WITH BLOCKERS
What exactly is barcode blocking?
It temporarily blocks the complementary adapter sequences of library fragments
from hybridizing
Barcode blockers are annealed to adapter sequences to prevent non-specific,
off-target pull down
Benefits of barcode blocking include...
Improving the number of reads on-target
Increasing the depth of enrichment
So what is the difference betweenAgilent’s and Bioo Scientific’s
barcode blocking strategy?
Agilent’s barcode blocking strategy...
Incorporates non-specific barcode blockers that work with all adapter sequences
On the other hand, Bioo Scientific’s barcode blocking strategy is index-specific
Index-specific barcode blockers are used & pooled depending upon the indexes used
during library prep
Why is using an index-specific barcode blocking strategy more beneficial?
Index-specific barcode blocking strategies improves blocking of adapter sequence
during hybridization
Index-specific barcode blocking strategies result in higher % of on-target reads
NEXTflex Agilent
Targeted Bases 10,180,000 10,180,000Reads Mapped 57,608,301 55,621,420
% Unique / Duplication 95.5 / 4.5 95.7 / 4.3% On Target 83.3 78.5
Mean Coverage 151.2 137.8
Index-specific barcode blocking strategies result in better coverage statistics
NEXTflex Agilent
Targeted Bases 10,180,000 10,180,000Reads Mapped 57,608,301 55,621,420
% Unique / Duplication 95.5 / 4.5 95.7 / 4.3% On Target 83.3 78.5
Mean Coverage 151.2 137.8
The NEXTflex™ Pre- & Post- Capture Combo Kit (Agilent SureSelectXT2 Compatible) incorporates
index-specific barcode blocking strategies
Offering higher % of on-target reads & better coverage for Agilent SureSelectXT2
target enrichment
With enhanced adapter ligation efficiency and fewer library prep steps
The NEXTflex™ Pre- & Post- Capture Combo Kit (Agilent SureSelectXT2 Compatible) includes:
NEXTflex™ Rapid DNA-Seq library prep reagentsIndexed barcodes
Index-specific blockersHybridization and wash buffers
Post-capture amplification reagents
Learn more about the NEXTflex™ Pre- & Post- Capture Combo Kit (Agilent
SureSelectXT2 Compatible)
PURCHASE KIT DOWNLOADWHITEPAPER
If you have any questions email us [email protected]
or visit our website atBiooScientific.com/SureSelect
