Technical data

4 Hybridization
Step 1. Pool indexed DNA samples for hybridization
56 SureSelect
XT2
Target Enrichment System for Illumina
Step 1. Pool indexed DNA samples for hybridization
In this step, you pool the indexed gDNA samples, before hybridization to
the SureSelect
XT2
Capture Library.
Each hybridization reaction requires a total of 1500 ng indexed gDNA,
made up of equal amounts of 8 or 16 individual libraries. See Table 23 for
the recommended pool composition for your SureSelect
XT2
Capture
Library.
1 For each capture reaction pool, combine the appropriate volume of each
indexed gDNA library sample in one well of a SureCycler 8800 PCR
plate.
Each final capture reaction pool should contain 1500 ng indexed gDNA.
2 Use a vacuum concentrator, held at  45°C, to reduce the volume in
each well to <7 µL.
Avoid completely drying the sample. Over-drying the indexed library
pool negatively impacts target enrichment.
3 Add sufficient nuclease-free water to each concentrated gDNA pool to
bring the final well volume to 7 µL.
4 Cap the wells, then vortex the plate vigorously for 30 seconds. Spin in
a centrifuge or mini-plate spinner to collect the liquid at the bottom of
the wells.
Table 23
Capture Library size Number of gDNA
libraries per pool
Amount of each
indexed library in pool
Custom Capture Libraries 16 93.75 ng
Human DNA Kinome 16 93.75 ng
Inherited Disease 8 187.5 ng
Human or Mouse All-Exon 8 187.5 ng