Technical data

5 Indexing and Sample Processing for Multiplexed Sequencing
Step 1B. Amplify the captured libraries with indexing primers containing 6-bp indexes 1–16
76 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
6 T
ransfer the PCR plate or strip tube to a thermal cycler and run the
PCR amplification program shown in Table 39.
Table 43 Post-Capture PCR cycling program
7 When the PCR amplification program is complete, spin the plate or
strip tube or strip tube briefly.
Proceed to “Step 2. Purify the amplified captured libraries using
AMPure XP beads” on page 77.
Segment Number of Cycles Temperature Time
1 1 98°C 2 minutes
2 10 to 16 Cycles
See Table 44 for recommendations based
on Capture Library size
98°C 30 seconds
57°C 30 seconds
72°C 1 minute
3 1 72°C 10 minutes
4 1 4°C Hold
Table 44 Post-capture PCR cycle number recommendations
Capture Size Cycles
1 kb up to 0.5 Mb 16 cycles
0.5 Mb up to 1.49 Mb 14 cycles
>1.5 Mb 12 cycles
All Exon and Exome libraries 10 to 12 cycles
OneSeq libraries (all designs) 10 cycles