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Old 01-18-2019, 09:35 AM   #1
mlai2567
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Location: New York

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Default RNA-Seq Sample Question

Hi all,

I am preparing samples to send for RNA-sequencing. I've transformed a majority of my samples to cDNA, keeping only a few microliters of highly concentrated RNA for QC analysis, as I had concerns over the RNA stability. However, on the sequencing service page, it seems as if most samples are sent in RNA form. Has anyone successfully sent cDNA samples for RNA-sequencing?

Thanks in advance,
Michael
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Old 01-18-2019, 11:58 AM   #2
BioDynami
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The commercial RNA-Seq kits are using RNA as input, not cDNA. You have converted RNA to cDNA, the single-stranded DNA? It may be a problem for the sequencing cores.
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Old 01-19-2019, 07:31 AM   #3
Bukowski
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Of course you can submit cDNA for sequencing, at this point it's just 'DNA'. Most cores would probably prefer to take the RNA because then the library preparation is in their hands, and not yours - and they can do the QC that follows the process to maximise their confidence in the seqeuncing of your samples.

You just need to speak to the provider and tell them what the DNA source is.

Next time, just send the RNA.
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Old 01-19-2019, 05:26 PM   #4
luc
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Bukowski is correct. RNA-seq library prep protocols (starting from RNA) are optimized for consistency to allow quantitative DGE analysis. Your cDNA prep protocol might not be.
Is your cDNA double-stranded (as required for most DNA library preps) or single-stranded?
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