Technical data

Sample Preparation (200 ng DNA Samples) 3
Step 5. Adenylate the 3' end of the DNA fragments
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 49
Step 5. Adenylate the 3' end of the DNA fragments
Use the SureSelect XT Library Prep Kit ILM for this step.
Hold samples on ice while setting up this step.
1 Prepare t
he appropriate volume of Adenylation master mix, as
described in Table 23, on ice. Mix well on a vortex mixer.
Table 23 Preparation of Adenylation master mix
2 Add 20 µL of the Adenylation master mix to each end-repaired,
purified DNA sample (approximately 30 µL).
3 Mix well by pipetting up and down.
4 Incubate the plate in the thermal cycler and run the program in
Table 24. Do not use a heated lid.
Table 24 dA-Tailing Thermal Cycler Program
Reagent Volume for 1 reaction Volume for 16 reactions
(includes excess)
Nuclease-free water 11 µL 181.5 µL
10× Klenow Polymerase Buffer (blue cap) 5 µL 82.5 µL
dATP (green cap) 1 µL 16.5 µL
Exo(–) Klenow (red cap) 3 µL 49.5 µL
Total 20 µL 330 µL
Step Temperature Time
Step 1 37°C 30 minutes
Step 2 4°C Hold