Technical data

Post-Capture Sample Processing for Multiplexed Sequencing 5
Step 1. Amplify the captured libraries
SureSelect
XT2
Target Enrichment System for Illumina 65
5 Place t
he plate in a thermal cycler and run the PCR amplification
program shown in Table 28 using the cycle number specified in
Table 29.
Table 28 Post-Capture PCR cycling program
Table 29 Recommended cycle number based on SureSelect Capture Library size
Amplify the captured DNA using a minimal number of PCR cycles. If yield is too low, or too
high (where non-specific high molecular weight products are observed), adjust the number
of cycles accordingly with the remaining captured DNA template.
Segment Number of
Cycles
Temperature Time
1 1 98°C 2 minutes
2 8-11
see Table 29
98°C 30 seconds
60°C 30 seconds
72°C 1 minute
3 1 72°C 10 minutes
4 1 4°C Hold
Size of SureSelect
Capture Library
Cycles
1 to 499 kb 12 to 14 cycles
0.5 to 1.49 Mb 9 to 11 cycles
> 1.5 Mb 8 to 10 cycles
All Exon 8 to 10 cycles
NOTE