Technical data

Before You Begin 1
Procedural Notes
SureSelect
XT2
Target Enrichment System for Illumina 13
Procedural Notes
• This User Guide includes protocols for library preparation using either
1 g DNA samples (see Chapter 2 on page 19) or 100 ng DNA samples
(see Chapter 3 on page 37). Make sure that you are following the
appropriate protocol for your DNA input amount. After the prepared
libraries are amplified, both DNA input options use the same protocol
for hybridization and post-capture processing.
• 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.
• Maintain a clean work area.
• Do not mix reactions containing gDNA on a vortex mixer. Instead,
gently tap the tube with your finger to mix the sample.
• Avoid repeated freeze-thaw cycles of stock and diluted gDNA solutions.
Possible stopping points, where gDNA samples may be stored overnight
at 4°C, are marked in the protocol. When storing samples for >24 hours,
store the samples at –20°C, but do not subject the samples to multiple
freeze/thaw cycles.
• When preparing 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 vials used during an experiment on ice or in a cold block.
4 Library Preparation Master Mixes should not be frozen and thawed
more than five times. If you plan to use the reagents in more than
five experiments, aliquot to multiple vials to minimize freeze/thaw
cycles for each vial.
• In general, follow Biosafety Level 1 (BL1) safety rules.
Safety Notes
• Wear appropriate personal protective equipment (PPE) when working in the
laboratory.
CAUTION