Technical data

4 Hybridization and Capture
Step 2. Prepare streptavidin-coated magnetic beads
66 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
Step 2. Prepare streptavidin-coated magnetic beads
The hybrid capture protocol uses reagents provided in SureSelect Target
Enrichment Box 1 (stored at room temperature) in addition to the
streptavidin-coated magnetic beads obtained from another supplier (see
Table 2 on page 14).
1 Pre
warm SureSelect Wash Buffer 2 at 65°C in a circulating water bath
or heat block for use in “Step 3. Capture the hybridized DNA using
streptavidin-coated beads” on page 67.
2 Vigorously resuspend the Dynabeads MyOne Streptavidin T1 magnetic
beads on a vortex mixer. The magnetic beads settle during storage.
3 For each hybridization sample, add 50 µL of the resuspended beads to
wells of a fresh PCR plate or strip tube.
4 Wash the beads:
a Add 200 µL of SureSelect Binding Buffer.
b Mix by pipetting up and down until beads are fully resuspended.
c Put the plate or strip tube into a magnetic separator device.
d Wait until the solution is clear, then remove and discard the
supernatant.
e Repeat step a through step d two more times for a total of 3 washes.
5 Resuspend the beads in 200 µL of SureSelect Binding Buffer.
If you are equipped for higher-volume magnetic bead captures, the streptavidin beads may
be batch-washed in an Eppendorf tube or conical vial. Start the batch-washing procedure
using excess bead solution. After resuspending the washed beads in the appropriate
volume of SureSelect Binding Buffer, aliquot 200 l of the washed beads to plate or strip
tube wells to be used for hybridization capture.
NOTE