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Dear all,
As the title suggests I am running a batch of small RNA sequencing using the SOLiD system. Having extracted good quality total RNA (RIN >9), the yield of some samples is rather low (as low as 20ng/ul). Therefore I'm trying to find out whether gDNA could possibly affect small RNA sequencing in an attempt to avoid this step and the likely sample loss.
My thinking is that I perform the ligation reaction (which favours single stranded species) and then following cDNA synthesis, size select to only include the miRNA size range. Given that gDNA is huge, and that I don't perform any sort of digest or sonication, I am right in thinking there is a limited chance of me including any gDNA in my library.
Happy to be completely wrong...!
D :-)
As the title suggests I am running a batch of small RNA sequencing using the SOLiD system. Having extracted good quality total RNA (RIN >9), the yield of some samples is rather low (as low as 20ng/ul). Therefore I'm trying to find out whether gDNA could possibly affect small RNA sequencing in an attempt to avoid this step and the likely sample loss.
My thinking is that I perform the ligation reaction (which favours single stranded species) and then following cDNA synthesis, size select to only include the miRNA size range. Given that gDNA is huge, and that I don't perform any sort of digest or sonication, I am right in thinking there is a limited chance of me including any gDNA in my library.
Happy to be completely wrong...!
D :-)
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