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Preparing samples for PacBio iso-seq sequencing

We utilise the template switching capability of certain reverse transcriptases to generate and amplify cDNA for whole transcript sequencing. To perform Iso-seq, you need to supply:

  • RNA with a poly-A tail (or ribosome deplete and then adenylate yourself for prokaryotic RNA)
  • ≥300 ng of Total RNA input recommended (although we can work with lower yields of RNA if integrity is good
  • RNA integrity number ≥7.0 (ideally ≥8.0)
    • Please check on an Agilent tapestation, bioanalyzer or equivalent
  • Ensure the RNA is dissolved in water free from contaminants
    • No phenol, no salts (can be detected by spectrophotometry/nanodrop)

Please contact us for information on iso-seq sequencing.