Thermo Scientific Applied Biosystems SeqStudio Genetic Analyzer - Prepare the Samples

Prepare the samples
Sample preparation guidelines
Item
Guidelines
Plates and tubes
• Use MicroAmp
™
Optical 96-Well Reaction Plate or MicroAmp
™
Reaction Tubes with
a tray and retainer set.
IMPORTANT! Fast plates are not compatible with the SeqStudio
™
Genetic
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.
Fragment and HID analysis
sample preparation
Intensity (RFU)
Size (bp)
• 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 DNA Fragment Analysis by Capillary Electrophoresis
User Guide (Pub. No. 4474504).
Sequence analysis sample
preparation
• 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
™
Formamide.
• Use a 65 µL or 130 µL sample volume for samples that are prepared with the
BigDye XTerminator
™
Purification Kit. See BigDye XTerminator
™
Purification Kit
Protocol (Pub. No. 4374408).
IMPORTANT! Use the appropriate run modules for samples prepared with
BigDye XTerminator
™
Purification Kit. See “Run modules, read lengths, size
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 DNA Sequencing by Capillary Electrophoresis
Chemistry Guide Second Edition (Pub. No. 4305080).
Chapter 2 Prepare the samples and the instrument
Prepare the samples
2
SeqStudio
™
Genetic Analyzer Instrument and Software User Guide
37