Single Cell mRNA Seq basic question
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6.4 years ago
CuriousSeq ▴ 20

If an average human somatic cell has approximately 12k transcripts of mRNA floating around, how do we (for example) get a mean reads per cell of 100k? Shouldn't you get 12k mean reads per cell

RNA-Seq • 1.1k views
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Number of reads you will get is very different than the number of transcripts due to shearing the DNA and PCR amplification.

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So are two reads with the same UMI considered two reads or one read?

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They are two reads but they originated from the same fragment.

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there are lots of other non-mRNA transcript floating around in the cell (and yes despite poly-A capture you will pick some of them up as well)

EDIT: ok, my bad, indeed number of reads is something very different than number of transcripts as goodez said.

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I have seen estimates of 500k transcripts. It will vary by cell type, though. And, as others have pointed out, one transcript can produce multiple reads.

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