Single stranded cDNA is used by race pcr and rt-pcr while next-gen sequencing (Illumina) requires double stranded cDNA. What is the reason behind this ?

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    $\begingroup$ I'm voting to close this question as off-topic because the question is not about bioinformatics, but for some specific methods of the wet lab. If you have a question about how to handle these sequences by computers, I think it would be better suited for the site. $\endgroup$ – llrs May 22 '18 at 13:27
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    $\begingroup$ Can we migrate this to biology? It's a valid question and I think it's even somewhat relevant to bioinformatic (it's important to understand generation of the data), but it would suit better to Biology.SE I think. Anyway, could you link source of your information? $\endgroup$ – Kamil S Jaron May 22 '18 at 14:41

Sequencing also requires single stranded DNA, otherwise the sequencing primers can’t bind. Sure there are steps in the protocol where double stranded material exists, but that’s transient.


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