Question: Does input rna concentration change expressions analysis?
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gravatar for badribio
29 days ago by
badribio240
badribio240 wrote:

Is there a relation between the quantity of rna/cdna that is used for library prep to number of reads that you seqeunce/quantify? for example: sample a and b are biological replicates 10ng cdna was used for sample a and 20ng for sample b will you see an increase in number of reads mapping to a given gene?

rna-seq • 108 views
ADD COMMENTlink modified 29 days ago by swbarnes25.5k • written 29 days ago by badribio240
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gravatar for swbarnes2
29 days ago by
swbarnes25.5k
United States
swbarnes25.5k wrote:

Assuming the quality is about the same, if you put in more than the minimum for that library protocol, it won't much matter much how much you put in. What matters more is how much library gets loaded on a flow cell, and what % of the lane/run it takes up.

ADD COMMENTlink modified 29 days ago • written 29 days ago by swbarnes25.5k

That makes sense, we ran in to a different problem when we plot the TPM's for each transcript between two different concentration (same biological replicate) there are some genes that have higher TPM in the sample with more input RNA.

ADD REPLYlink written 29 days ago by badribio240
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