Technical data
Indexing and Sample Processing for Multiplexed Sequencing 5
Step 2. Purify the amplified captured libraries using AMPure XP beads
SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing 77
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 fresh 70% ethanol per sample, plus excess, for use in
step 9.
3 Mix the AMPure XP bead suspension well so that the suspension
appears homogeneous and consistent in color.
4 Add 90 µL of the homogeneous AMPure XP bead suspension to each
50-µL amplified DNA sample bead suspension in the PCR plate or strip
tube.
5 Mix thoroughly by pipetting up and down.
Check that the beads are in a homogeneous suspension in the sample
wells. Each well should have a uniform color with no layers of beads or
clear liquid present.
6 Incubate samples for 5 minutes at room temperature.
7 Put the plate or strip tube on the magnetic stand at room temperature.
Wait for the solution to clear (approximately 3 to 5 minutes).
8 While keeping the plate or tubes in the magnetic stand, carefully
remove and discard the cleared solution from each well. Do not disturb
the beads while removing the solution.
9 Continue to keep the plate or tubes in the magnetic stand while you
dispense 200 µL of freshly prepared 70% ethanol in each sample well.
10 Wait for 1 minute to allow any disturbed beads to settle, then remove
the ethanol.
11 Repeat step 9 and step 10 once for a total of two washes. Make sure to
remove all of the ethanol at each wash step.
12 Seal the wells with strip caps, then briefly spin to collect the residual
ethanol. Return the plate or strip tube to the magnetic stand for
30 seconds. Remove the residual ethanol with a P20 pipette.
13 Dry the samples by placing the unsealed plate or strip tube on the
thermal cycler, set to hold samples at 37°C, for 1 to 2 minutes or until
the residual ethanol completely evaporates.
14 Add 30 µL of nuclease-free water to each sample well.