Technical data

4 Hybridization
Step 4. Capture the hybridized DNA using streptavidin beads
60 SureSelect
XT2
Target Enrichment System for Illumina
Step 4. Capture the hybridized DNA using streptavidin beads
1 Estimate and record the volume of hybridization solution that remains
after the 24 hour incubation.
2 Maintain the hybridization plate at 65°C while you use a multichannel
pipette to transfer the entire volume (approximately 60 µL) of each
hybridization mixture to the plate wells containing 200 µL of washed
streptavidin beads.
Mix well by slowly pipetting up and down 3 to 5 times.
3 Cap the wells, then incubate the capture plate on a Nutator mixer or
equivalent for 30 minutes at room temperature.
Make sure the samples are properly mixing in the wells.
4 Briefly spin the plate in a centrifuge or mini-plate spinner.
5 Put the plate in a magnetic separator to collect the beads from the
suspension. Remove and discard the supernatant.
6 Resuspend the beads in 200 µL of SureSelect XT2 Wash 1. Mix by
pipetting up and down until the beads are fully resuspended.
7 Briefly spin in a centrifuge or mini-plate spinner.
8 Put the plate in the magnetic separator.
9 Wait for the solution to clear, then remove and discard the supernatant.
CAUTION
Excessive evaporation, such as when less than 52 µL remains after hybridization, can
indicate suboptimal capture performance.
CAUTION
It is important to maintain bead suspensions at 65°C during the washing procedure
below to ensure specificity of capture.
Make sure that the SureSelect XT2 Wash 2 is pre-warmed to 65°C before use.
Do not use a tissue incubator, or other devices with significant temperature
fluctuations, for the incubation steps.