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Sample Requirements


NGS - General Sample Requirements

MiSeq®/NextSeq® Platforms

DNA samples for MiSeq®/NextSeq®

  • 1-2 µg of high-quality DNA with a concentration above 10 ng/µL, determined by fluorometry or by electrophoresis on a 1% agarose gel using a 1Kb Molecular Marker, are required for general library preparation protocols.
  • DNA must be double-stranded, integrate, RNA and inhibitor-free. We recommend checking the DNA quality by electrophoresis on a 1% agarose gel alongside a 1Kb Molecular Marker
  • OD260/280 ratio, determined by spectrophotometry (Nanodrop System, for example), should be between 1.8 and 1.95.
  • DNA samples should be sent in10 mMTris-HCl, pH 8.0 buffer.

Please send an electrophoresis image of an 80 ng sample run on a 1% agarose gel with the appropriate molecular marker.

RNA samples for MiSeq®/NextSeq®

  • 1-4 µg of total RNA, determined by fluorometry, are required for general library preparation protocols.
  • We recommend RNA concentration be determined by fluorometric assays. The Qubit RNA HS Assay Kit may be used for this quantification.
  • The RNA integrity should be verified with the RNA Pico 6000 Assay Kit on the Agilent 2100 Bioanalyzer. Good results are obtained when the RNA Integrity Number (RIN) is above 8.
  • RNA samples should be DNase treated to remove DNA contaminants.
  • OD260/280 andOD260/230 ratios, determined by spectroscopy (Nanodrop System, for example), should be >1.8.
  • RNA samples can be sent in Nuclease-Free water or stabilization solution (RNAlater) and shipped on dry ice.

Samples for amplicon sequencing or microbial profiling

  • For microbial profiling studies and to guarantee the presence of DNA and absence of contaminants, samples must be previously checked by PCR amplification. Please send a minimum volume of 20 µl, per sample.
  • For sequencing of other amplicons, DNA samples with a concentration ≥ 50 ng/µL and the OD260/280ratio between 1.75 and 1.95 are required. The DNA quantity sent will depend on the number of fragments to be sequenced (100 ng per amplicon).

Samples are checked and quality controlled upon arrival. If samples fail the QC procedures (quality and quantity), a new sample will be requested.

Samples must be sent clearly identified and in accordance with the sample submission form. Please send all quality control procedures applied to the samples.

If you have any other question or need additional information regarding samples or quality control procedures, please contact our services.

 

ION PROTON™ Platform

DNA Samples for Ion Proton™

  • 2-3 µg of high-quality DNA with an approximate concentration of 100 ng/µL, determined by fluorometry, are required for general library preparation protocols. The Qubit ds DNA HS Assay Kit can be used for the fluorescence quantification.
  • DNA must be double-stranded, integrate, RNA and inhibitor-free. We recommend checking the DNA quality by electrophoresis on a 1% agarose gel alongside a 1Kb Molecular Marker.
  • OD260/280 andOD260/230  ratios, determined by spectrophotometry (Nanodrop System, for example), should be between 1.75 and 1.95.
  • DNA samples should be sent in10 mMTris-HCl, pH 8.0 buffer.

Please send an electrophoresis image of an 80 ng sample run on a 1% agarose gel with the appropriate molecular marker.


RNA Samples for Ion Proton

  • 50 µg of total RNA or 1 µg Poly(A) RNA or 1 µg ribosomal depleted RNA, determined by fluorometry, are required for general library preparation protocols.
  • The preparation of libraries using the Ion AmpliSeq™ RNA technology requires 1 µg of high-quality total RNA.
  • We recommend RNA concentration be determined by fluorometric assays. The Qubit RNA HS Assay Kit may be used for this quantification.
  • The RNA integrity should be verified with the RNA Pico 6000 Assay Kit on the Agilent 2100 Bioanalyzer. Good results are obtained when the RNA Integrity Number (RIN) is above 8.
  • RNA samples should be DNase treated to remove DNA contaminants.
  • OD260/280 andOD260/230  ratios, determined by spectrophotometry (Nanodrop System, for example), should be >1.8.
  • RNA samples can be sent in Nuclease-Free water or stabilization solution (RNAlater) and shipped on dry ice.

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