Technical data

3 Sample Preparation (100 ng DNA Samples)
Step 8. Amplify the indexed library
50 SureSelect
XT2
Target Enrichment System for Illumina
Step 8. Amplify the indexed library
1 Prepare the appropriate volume of pre-capture PCR reaction mix, as
described in Table 21, on ice. Mix well on a vortex mixer.
2 In separate wells of a SureCycler 8800 PCR plate, combine 26 µL of the
amplification mixture prepared in Table 21 and 24 µL of each indexed
gDNA library sample. Mix by pipetting. Change pipette tips between
samples.
3 Run the program in Table 22 in a SureCycler thermal cycler.
CAUTION
To avoid cross-contaminating libraries, set up PCR reactions (all components except
the library DNA) in a dedicated clean area or PCR hood with UV sterilization and
positive air flow.
Table 21 Preparation of SureSelect Pre-Capture PCR Reaction Mix
Reagent Volume for 1
reaction
Volume for 16 reactions
(includes excess)
XT2 Primer Mix 1 µL 16.5 µL
Herculase II PCR Master Mix 25 µL 412.5 µL
Total 26 µL 429 µL
Table 22 Pre-Capture PCR Thermal Cycler Program
Segment Number of Cycles Temperature Time
1 1 98°C 2 minutes
2 8 98°C 30 seconds
60°C 30 seconds
72°C 1 minute
3 1 72°C 10 minutes
4 1 4°C Hold