SureSelectXT2 Target Capture is a solution-based hybridization system that enriches genomic regions of interest rather than whole genomes. It uses a protocol involving DNA isolation, library preparation with barcodes, hybridization with blockers, hybridization of SureSelect libraries, washing and recovering captured DNA, amplification of captured DNA, and quantitation. Barcode blocking temporarily blocks adapter sequences to prevent non-specific binding. Bioo Scientific's index-specific barcode blocking strategy improves blocking and results in a higher percentage of on-target reads and better coverage statistics compared to Agilent's non-specific strategy. The NEXTflex kit from Bioo Scientific incorporates this index-specific approach.

1. Improving the Performance of
SureSelectXT2
Target Capture

2. What is SureSelectXT2
Target Capture?

3. A solution-based hybridization
target capture system that enriches
for regions of interest, as opposed to
whole-genome sequencing

4. A wide range of predesigned probe
sets or baits and flexible customization
options allows users to design their
own capture panels

5. To see how it works, next we’ll take a look at
the target capture protocol

6. 0101 DNA ISOLATION

7. 01
02
01
02
DNA ISOLATION
LIBRARY PREP WITH BARCODES

8. 01
02
03
01
02
03
DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS

9. 01
02
04
03
01
02
04
03
DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES

10. 01
02
04
03
05
01
02
04
03
05
DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
WASH & RECOVER CAPTURED DNA

11. 01
02
04
06
03
05
01
02
04
06
03
05
DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
AMPLIFY CAPTURED DNA
WASH & RECOVER CAPTURED DNA

12. 01
02
04
06
03
05
07
01
02
04
06
03
05
07
DNA ISOLATION
LIBRARY PREP WITH BARCODES
HYBRIDIZE WITH BLOCKERS
HYBRIDIZE SURESELECT LIBRARIES
AMPLIFY CAPTURED DNA
WASH & RECOVER CAPTURED DNA
QUANTITATE USING qPCR
ContainedinBiooScientific’sKit

13. Now that we’ve seen the protocol, let’s focus
on Step 3: HYBRIDIZE WITH BLOCKERS
0303 HYBRIDIZE WITH BLOCKERS

14. What exactly is barcode blocking?

15. It temporarily blocks the complementary
adapter sequences of library fragments
from hybridizing

16. Barcode blockers are annealed to adapter
sequences to prevent non-specific,
off-target pull down

17. Benefits of barcode blocking include...

18. Improving the number of reads on-target

19. Increasing the depth of enrichment

20. So what is the difference between
Agilent’s and Bioo Scientific’s
barcode blocking strategy?

21. Agilent’s barcode blocking strategy...

22. Incorporates non-specific barcode blockers
that work with all adapter sequences

23. On the other hand, Bioo Scientific’s barcode
blocking strategy is index-specific

24. Index-specific barcode blockers are used &
pooled depending upon the indexes used
during library prep

25. Why is using an index-specific barcode
blocking strategy more beneficial?

26. Index-specific barcode blocking strategies
improves blocking of adapter sequence
during hybridization

27. Index-specific barcode blocking strategies
result in higher % of on-target reads
NEXTflex Agilent
Targeted Bases 10,180,000 10,180,000
Reads 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

28. Index-specific barcode blocking strategies
result in better coverage statistics
NEXTflex Agilent
Targeted Bases 10,180,000 10,180,000
Reads 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

29. The NEXTflex™ Pre- & Post- Capture Combo Kit
(Agilent SureSelectXT2
Compatible) incorporates
index-specific barcode blocking strategies

30. Offering higher % of on-target reads &
better coverage for Agilent SureSelectXT2
target enrichment

31. With enhanced adapter ligation efficiency
and fewer library prep steps

32. The NEXTflex™ Pre- & Post- Capture Combo Kit
(Agilent SureSelectXT2
Compatible) includes:
NEXTflex™ Rapid DNA-Seq library prep reagents
Indexed barcodes
Index-specific blockers
Hybridization and wash buffers
Post-capture amplification reagents

33. Learn more about the NEXTflex™ Pre-
& Post- Capture Combo Kit (Agilent
SureSelectXT2
Compatible)
PURCHASE KIT DOWNLOAD
WHITEPAPER

34. If you have any questions email us at
nextgen@biooscientific.com
or visit our website at
BiooScientific.com/SureSelect