Technical data

5 Indexing and Sample Processing for Multiplexed Sequencing
Step 1A. Amplify the captured libraries with indexing primers containing 8-bp indexes A01–H12
72 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
5 A
dd the DNA library samples to the PCR reactions:
a Obt
ain the PCR plate or strip tube containing 30 µL of bead-bound
target-enriched DNA samples from ice (prepared on page 68).
b Pipett
e each DNA sample up and down until the bead suspension is
homogeneous, then transfer 14 µL of the sample to the appropriate
well of the PCR plate or strip tube containing PCR reaction mix and
indexing primer.
c Mix t
he PCR reactions well by pipetting.
d Store the remaining library-bound beads at –20°C for future use, if
needed.
6 Transfer the PCR plate or strip tube to a thermal cycler and run the
PCR amplification program shown in Table 39.
Table 39 Post-Capture PCR cycling program
Segment Number of Cycles Temperature Time
1 1 98°C 2 minutes
2 10 to 16 Cycles
See Table 40 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