Technical data

Sample Preparation (3 µg DNA Samples) 2
Step 10. Amplify the adaptor-ligated library
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 35
1 Prepare the appropriate volume of pre-capture PCR reaction mix, as
described in Table 17, on ice. Mix well on a vortex mixer.
2 Combine 35 µL of the PCR reaction mixture prepared in Table 17 and
15 µL of each purified DNA library sample from step 16 on page 33.
Add a single DNA library sample to each well of the plate or strip tube.
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 17 Preparation of SureSelect Pre-Capture PCR Reaction Mix
Reagent Volume for 1
reaction
Volume for 16 reactions
(includes excess)
Nuclease-free water 21 µL 346.5 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 8.25 µL
Herculase II Fusion DNA Polymerase (red cap) 1 µL 16.5 µL
Total 35 µL 577.5 µL