Technical data

Hybridization and Capture 4
Step 1. Hybridize DNA samples to the Capture Library
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 63
6 To each gDNA library sample well prepared in step 3 on page 61, add
5.6 µL of the SureSelect Block Mix prepared in Table 32. Pipette up
and down to mix.
7 Cap the wells, then transfer the sealed plate or strip tube to the
thermal cycler and run the following program shown in Table 33.
Use a heated lid, set at 105°C, to hold the temperature at 65°C.
Make sure that the DNA + Block Mix samples are held at 65°C for at
least 5 minutes before adding the remaining hybridization reaction
components in step 10 below.
Table 33 Thermal cycler program for DNA + Block Mix prior to hybridization
8 Prepare the appropriate dilution of SureSelect RNase Block, based on
the size of your Capture Library, according to Table 34. Prepare the
amount required for the number of hybridization reactions in the run,
plus excess.
Table 34 Preparation of RNase Block dilution
Keep the mixture on ice until it is used in step 9.
CAUTION
For each protocol step that requires removal of tube cap strips, make sure to reseal the
tubes with a fresh strip of caps. Reuse of strip caps can cause sample loss, sample
contamination, or imprecision in sample temperatures during incubations.
Step Temperature Time
Step 1 95°C 5 minutes
Step 2 65°C Hold (at least 5 minutes)
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 25% (1:3) 2 µL
<3.0 Mb 10% (1:9) 5 µL