Thermo Scientific Ion Torrent A45727, Руководство пользователя

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Dilute the samples and load the
sample plate
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Guidelines for nucleic acid isolation and quantification—sample runs . . . . . . . 68
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Dilute the samples (if needed) and load the sample plate—sample run . . . . . . 69
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Guidelines for library quantification—library runs . . . . . . . . . . . . . . . . . . . . . . . . 71
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Dilute and pool libraries, and load the sample plate—library run . . . . . . . . . . . 71
Before starting a run on the instrument, you must quantify and dilute the samples or
sample libraries, then load the sample plate.
Guidelines for nucleic acid isolation and quantification—sample
runs
These are general guidelines for isolating and quantifying DNA and RNA. For assay-
specific guidelines, see the assay user guide.
• See “Recommended materials for nucleic acid isolation and quantification“ on
page 18 for recommended kits for isolating DNA and RNA.
• We recommend the TaqMan ™ RNase P Detection Reagents Kit (Cat. No. 4316831)
for quantifying amplifiable human genomic DNA (see Demonstrated Protocol:
Sample Quantification for Ion AmpliSeq ™ Library Preparation Using the TaqMan ™
RNAse P Detection Reagents Kit (Pub. No. MAN0007732) available at
thermofisher.com).
• The Qubit ™ dsDNA HS Assay Kit (Cat. No. Q32851 or Q32854) can also be used
for quantification, particularly for FFPE DNA, and highly degraded DNA
samples. See “Quantify FFPE DNA with the Qubit
™
Fluorometer“ on page 135 for
a detailed procedure for quantifying FFPE DNA.
• We recommend the Qubit
™
RNA HS Assay Kit (Cat. No. Q32855) for quantifying
RNA.
• Quantification methods such as densitometry (for example, using a NanoDrop
™
spectrophotometer) are not recommended, because they are not specific for DNA
or RNA. Use of these methods can lead to gross overestimation of the
concentration of sample nucleic acid, under-seeding of the target amplification
reaction, and low library yields.
• The Ion AmpliSeq ™ Direct FFPE DNA Kit bypasses nucleic acid isolation when
preparing libraries from FFPE sections on slides. See the Ion AmpliSeq ™ Direct
FFPE DNA Kit User Guide (Pub. No. MAN0014881) for a protocol for using this kit
to prepare gDNA from FFPE tissue.
• The Direct FFPE DNA preparation can be stored for up to 6 months at –30°C to
−10°C before library preparation.
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68 Genexus
™
Integrated Sequencer User Guide