RNA-seq library size difference

foxytomato

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Join Date: Oct 2009

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#1

RNA-seq library size difference

09-17-2013, 01:31 PM

Hello,

I have 4 samples of RNA-seq library made with scriptseq kit.
Bioanalyzer shows that for one sample the peak size is about 30 bp smaller than others and the size distribution is overall shifted too.
(270 bp for this one and 300 bp for others)
Would this make a huge problem when I analyze the final result?
Or all 4 libraries should be made again?
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zatoichi888

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Join Date: May 2013

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#2

09-19-2013, 07:07 AM

I'm relatively new to RNAseq but have done a few times library prep. If you start with same amount of input RNA saying 20ng of mRNA(either rRNA depleted or polyA purification) and downstream treatments are exactly the same, you ought be able to get similar average library size. I got from 290 to 310bp average size for my 12 samples at one batch.
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