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Old 06-05-2012, 06:30 PM   #1
luc
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Location: US

Join Date: Dec 2010
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Default How much cDNA out of RNA? - RNAseq library prep

When generating RNAseq libraries (using RNA fragmentation, the "standard" reverse transcription with , Superscript III, random hexamers, and second strand synthesis with DNA Polymerase I/RNase H) - how much DNA would one expect to generate from about 100 ng mRNA?

We have been getting about 50 ng DNA (on average 250 bp fragments, before adapter ligation) - is this amount in the expected range?

Thanks a lot in advance.
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