Technical data

2 Sample Preparation (3 µg DNA Samples)
Step 8. Ligate the paired-end adaptor
32 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
Step 8. Ligate the paired-end adaptor
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 Ligation master mix, as described in
Table 14, on ice. Mix well on a vortex mixer.
Table 14 Preparation of Ligation master mix
2 Add 37 µL of the Ligation master mix to each dA-tailed, purified DNA
sample (13 µL) in the PCR plate wells.
3 Mix well by pipetting up and down.
4 Incubate the plate in the thermal cycler and run the program in
Table 15. Do not use a heated lid.
Table 15 Ligation Thermal Cycler Program
Stopping Point If you do not continue to the next step, seal the plate and store at –20°C.
Reagent Volume for 1 reaction Volume for 16 reactions
(includes excess)
Nuclease-free water 15.5 µL 255.75 µL
5× T4 DNA Ligase Buffer (green cap) 10 µL 165 µL
SureSelect Adaptor Oligo Mix (brown cap) 10 µL 165 µL
T4 DNA Ligase (red cap) 1.5 µL 24.75 µL
Total 37 µL 610.5 µL
Step Temperature Time
Step 1 20°C 15 minutes
Step 2 4°C Hold