Technical data

5 Post-Capture Sample Processing for Multiplexed Sequencing
Step 2. Purify the amplified captured libraries using AMPure XP beads
66 SureSelect
XT2
Target Enrichment System for Illumina
Step 2. Purify the amplified captured libraries using
AMPure XP beads
1 Let the AMPure XP beads come to room temperature for at least
30
minutes. Do not freeze the beads at any time.
2 Prepare 400 µL of 70% ethanol per sample, plus excess, for use in
step 9.
3 Mix the bead suspension well so that the reagent appears homogeneous
and consistent in color.
4 Add 90 µL of the homogeneous AMPure XP bead suspension to each
sample well of the PCR plate, containing the 50-µL amplified captured
library samples (also containing streptavidin beads used for capture).
5 Mix well on a vortex mixer. Briefly spin the plate in a centrifuge or
mini-plate spinner to collect the liquid.
6 Incubate for 5 minutes at room temperature.
7 Put the plate into a magnetic separation device. Wait for the solution to
clear (approximately 3 to 5 minutes).
8 Keep the plate in the magnetic stand. Carefully remove and discard the
cleared solution from each well. Do not touch the beads while removing
the solution.
9 Continue to keep the plate in the magnetic stand while you dispense
200 µL of 70% ethanol into each sample well.
Use fresh 70% ethanol for optimal results.
10 Wait for 1 minute to allow any disturbed beads to settle, then remove
the ethanol.
11 Repeat step 9 and step 10 step once.
12 Seal the wells with strip caps, then briefly spin the plate to collect the
residual ethanol. Return the plate to the magnetic stand for 30 seconds.
Remove the residual ethanol with a P20 pipette.
13 Dry the samples on the SureCycler thermal cycler, set to hold samples
at 37°C, for 5 minutes or until the residual ethanol completely
evaporates.
14 Add 30 µL nuclease-free water to each sample well then mix well on a
vortex mixer. Briefly spin the plate in a centrifuge or mini-plate
spinner to collect the liquid.
15 Incubate for 2 minutes at room temperature.