Technical data

Hybridization and Capture 4
Step 3. Capture the hybridized DNA using streptavidin-coated beads
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 67
Step 3. Capture the hybridized DNA using streptavidin-coated
beads
1 Estimate and record the volume of hybridization solution that remains
after the 24 hour incubation.
2 Maint
ain the hybridization plate or strip tube at 65°C while you use a
multichannel pipette to transfer the entire volume (approximately 25 to
29 µL) of each hybridization mixture to the plate or strip tube wells
containing 200 µL of washed streptavidin beads.
Mix well by slowly pipetting up and down until beads are fully
resuspended.
Excessive evaporation, such as when less than 20 µL remains after hybridization, can
indicate suboptimal capture performance. SeeTable 58
on page 97 for tips to minimize
evaporation
.
3 Cap the wells, then incubate the capture plate or strip tube 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 or strip tube in a centrifuge or mini-plate
spinner.
5 Put the plate or strip tube in a magnetic separator to collect the beads.
Wait until the solution is clear, then remove and discard the
supernatant.
6 Resuspend the beads in 200 µL of SureSelect Wash Buffer 1. Mix by
pipetting up and down until beads are fully resuspended.
7 Incubate the samples for 15 minutes at room temperature.
8 Briefly spin in a centrifuge or mini-plate spinner.
9 Put the plate or strip tube in the magnetic separator. Wait for the
solution to clear, then remove and discard the supernatant.
NOTE