Prepare The Samples; Sample Preparation Guidelines - Thermo Scientific Applied Biosystems SeqStudio Genetic Analyzer User Manual

Prepare the samples

Sample preparation guidelines

Item
Plates and tubes
Fragment and HID analysis
sample preparation
Size (bp)
Sequence analysis sample
preparation
SeqStudio Genetic Analyzer Instrument and Software User Guide
™
• Use MicroAmp ™ Optical 96-Well Reaction Plate or MicroAmp
a tray and retainer set.
IMPORTANT! Fast plates are not compatible with the SeqStudio
Analyzer. Fast plates will damage the cartridge.
• Use the appropriate septa for plates and tubes.
• See "Required materials not supplied" on page 220 for more information.
• Prepare the samples as recommended by the kit for fragment analysis.
• Use a 10–20 µL sample volume.
• Ensure that Hi‑Di ™ Formamide is fresh.
– Hi‑Di ™ Formamide should not undergo more than two freeze-thaw cycles (one
to aliquot and one for use).
– Use the same day after thawing.
• For more information, see
User Guide (Pub. No. 4474504).
• Prepare sequencing reactions according to kit instructions, and purify the
extension products with ethanol precipitation, spin columns, or the BigDye
XTerminator ™ Purification Kit.
– If ethanol precipitation or spin columns are used, dry the samples in a
vacuum centrifuge without heat or at low heat for 10–15 minutes or until dry.
Note: Do not over dry the samples.
– Resuspend in 10–20 µL of Hi‑Di
• Use a 65 µL or 130 µL sample volume for samples that are prepared with the
BigDye XTerminator ™
Protocol (Pub. No. 4374408).
IMPORTANT! Use the appropriate run modules for samples prepared with
BigDye XTerminator ™
ranges, and run times" on page 140.
• Use a 10–20 µL sample volume for samples that are prepared with Hi‑Di
Formamide.
• Ensure that Hi‑Di ™ Formamide is fresh.
– Hi‑Di ™ Formamide should not undergo more than two freeze-thaw cycles (one
to aliquot and one for use).
– Use the same day after thawing.
• Do not resuspend samples in water, which can decrease sample stability.
• For more information, see
Chemistry Guide Second Edition (Pub. No. 4305080).
Chapter 2 Prepare the samples and the instrument
Guidelines
DNA Fragment Analysis by Capillary Electrophoresis
™ Formamide.
BigDye XTerminator
Purification Kit. See
Purification Kit. See "Run modules, read lengths, size
DNA Sequencing by Capillary Electrophoresis
2

Prepare the samples

™ Reaction Tubes with
™ Genetic
Purification Kit
™
™
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