Technical data

3 Sample Preparation (200 ng DNA Samples)
Step 9. Amplify the adaptor-ligated library
54 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
1 Prepare the appropriate volume of pre-capture PCR reaction mix, as
described in Table 28, on ice. Mix well on a vortex mixer.
2 Add 20 µL of the PCR reaction mixture prepared in Table 28 to each
purified DNA library sample (30 µL) in the PCR plate wells.
Mix by pipetting.
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 28 Preparation of SureSelect Pre-Capture PCR Reaction Mix
Reagent Volume for 1
reaction
Volume for 16 reactions
(includes excess)
Nuclease-free water 6 µL 99.0 µL
SureSelect Primer (brown cap) 1.25 µL 20.6 µL
SureSelect ILM Indexing Pre-Capture PCR
Reverse Primer (clear cap)
1.25 µL 20.6 µL
5× Herculase II Reaction Buffer (clear cap) 10 µL 165 µL
100 mM dNTP Mix (green cap) 0.5 µL 8.3 µL
Herculase II Fusion DNA Polymerase (red cap) 1 µL 16.5 µL
Total 20 µL 330 µL