Preparing samples for long read RNA or cDNA sequencing
We utilise the template switching capability of certain reverse transcriptases to generate and amplify cDNA for whole transcript sequencing. To perform long read RNAseq, you need to supply:
- RNA with a poly-A tail (or ribosome deplete and then adenylate yourself for prokaryotic RNA)
- ≥100 ng of polyA-RNA input recommended or 70-200ng already prepared cDNA
- 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)
For cDNA-PCR amplified libraries 1ng polyA-RNA or 50 ng total RNA
For Direct RNAsequencing 500ng polyA RNA in 9ul
Please contact us for information on ONT RNA/cDNA sequencing.