We have degraded RNA without any poly A tail from E. coli. We want to convert these RNA to cDNA and want to see if the strand has actually been synthesized? Size of these RNA ranges between 17-200 bases.
The only way to do this is doing a PCR over the cDNA target. If you thought something like Bioanalyzer or any other eletrophoresis similar approach, you will find a lot of potential artifacts that surely will mask your target.
Yes, but PCR needs ds cDNA for which we need to add adapter to the first strand of DNA (synthesized in reverse transcription). We need only first strand of the cDNA which will then be used in sequencing.
IMHO if you made in in large quantities, treated with RNase, you could detect some ssDNA.
I beleive Bioanalyzer can do the job for you if you have positive control (good quality RNA), negative control ( known degraded RNA) and your samples for compariosn.
I dont think so but your RIN number ya RNA integrety number if its above 7 (use bioanalyser) its bound to give good cDNA capturing all transcripts
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