Technical data

Sample Preparation (200 ng DNA Samples) 3
Step 7. Ligate the paired-end adaptor
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 51
Step 7. 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 Dilut
e the SureSelect Adaptor Oligo Mix (brown cap) 1:10 in
nuclease-free water immediately before use. Use the diluted oligo mix
when preparing the Ligation master mix in the next step.
2 Prepare the appropriate volume of Ligation master mix, as described in
Table 25, on ice. Mix well on a vortex mixer.
Table 25 Preparation of Ligation master mix
3 Add 37 µL of the Ligation master mix to each dA-tailed, purified DNA
sample (13 µL) in the PCR plate wells.
4 Mix well by pipetting up and down.
5 Incubate the plate in the thermal cycler and run the program in
Table 26. Do not use a heated lid.
Table 26 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
Diluted SureSelect Adaptor Oligo Mix from step 110 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