Technical data

1 Before You Begin
Procedural Notes
12 SureSelect
XT
Target Enrichment System for Illumina Multiplexed Sequencing
Procedural Notes
• To prevent contamination of reagents by nucleases, always wear
powder-free laboratory gloves and use dedicated solutions and pipettors
with nuclease-free aerosol-resistant tips.
• Use best-practices to prevent PCR product contamination of samples
throughout the workflow:
1 Assign separate pre-PCR and post-PCR work areas and use
dedicated equipment, supplies, and reagents in each area. In
particular, never use materials designated to post-PCR work areas for
pre-PCR segments of the workflow.
2 Maintain clean work areas. Clean pre-PCR surfaces that pose the
highest risk of contamination daily using a 10% bleach solution.
3 Always use dedicated pre-PCR pipettors with nuclease-free
aerosol-resistant tips to pipette dedicated pre-PCR solutions.
4 Wear powder-free gloves. Use good laboratory hygiene, including
changing gloves after contact with any potentially-contaminated
surfaces.
• Do not mix stock solutions of gDNA on a vortex mixer. Instead, gently
tap the tube with your finger to mix the sample.
• When preparing frozen reagent stock solutions for use:
1 Thaw the aliquot as rapidly as possible without heating above room
temperature.
2 Mix briefly on a vortex mixer, then spin in a centrifuge for 5 to
10 seconds to drive the contents off of walls and lid.
3 Store on ice or in a cold block until use.
• For each protocol step that requires removal of tube cap strips, reseal
the tubes with a fresh strip of domed caps. Cap deformation may result
from exposure of the cap strips to the heated lid of the thermal cycler
and from other procedural steps. Reuse of strip caps can cause sample
loss, sample contamination, or imprecision in sample temperatures
during thermal cycler incubation steps.
• In general, follow Biosafety Level 1 (BL1) safety rules.
• Possible stopping points, where samples may be stored at –20°C, are
marked in the protocol. Do not subject the samples to multiple
freeze/thaw cycles.