Technical data

Hybridization 4
Step 2. Hybridize gDNA library pools to the SureSelect Capture Library
SureSelect
XT2
Target Enrichment System for Illumina 57
Step 2. Hybridize gDNA library pools to the SureSelect
Capture Library
1 To each 7-µL indexed gDNA pool, add 9 µL of SureSelect XT2 Blocking
Mix. Pipette up and down to mix.
2 Cap the wells, then transfer the sealed plate to the thermal cycler and
run the following program shown in Table 24.
Use a heated lid, set at 105°C, to hold the temperature at 65°C.
Make sure that the plate is held at 65°C for at least 5 minutes before
the gDNA library/Block mixtures are used in step 7 below.
Table 24 Thermal cycler program used for sample denaturation prior to hybridization
3 Prepare the appropriate dilution of SureSelect RNase Block, based on
the size of your capture library, according to Table 25. Prepare the
amount required for the number of hybridization reactions in the run,
plus excess.
Table 25 Preparation of RNase Block dilution
Step Temperature Time
Step 1 95°C 5 minutes
Step 2 65°C Hold
CAUTION
The lid of the thermal cycler is hot and can cause burns. Use caution when working
near the lid.
Capture Library Size RNase Block dilution
(parts RNase Block:parts water)
Volume of dilute RNase Block
Required per hybridization reaction
<3.0 Mb 10% (1:9) 5 µL
>3.0 Mb 25% (1:3) 2 µL