Technical data

2 Sample Preparation (3 µg DNA Samples)
Step 4. Repair the ends
28 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
Step 4. Repair the ends
Use the SureSelect XT Library Prep Kit ILM for this step.
To process multiple samples, prepare master mixes with overage at each
step, without the DNA sample. Master mixes for preparation of 16 samples
(including excess) are shown in each table as an example.
Hold samples on ice while setting up this step.
1 Prepare the appropriate volume of End Repair master mix, as described
in Table 10, on ice. Mix well on a vortex mixer.
Table 10 Preparation of End Repair master mix
2 Add 52 µL of the master mix to each PCR plate sample well containing
purified, sheared DNA. Mix by pipetting up and down.
3 Incubate the plate in the thermal cycler and run the program in
Table 11. Do not use a heated lid.
Table 11 End-Repair Thermal Cycler Program
Reagent Volume for 1 reaction Volume for 16 reactions
(includes excess)
Nuclease-free water 35.2 µL 580.8 µL
10× End Repair Buffer (clear cap) 10 µL 165 µL
dNTP Mix (green cap) 1.6 µL 26.4 µL
T4 DNA Polymerase (purple cap) 1 µL 16.5 µL
Klenow DNA Polymerase (yellow cap) 2 µL 33 µL
T4 Polynucleotide Kinase (orange cap) 2.2 µL 36.3 µL
Total 52 µL 858 µL
Step Temperature Time
Step 1 20°C 30 minutes
Step 2 4°C Hold