ScriptSeq - 2nd peak @ 550bp

FerdinandvM

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Join Date: Jul 2013

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#1

ScriptSeq - 2nd peak @ 550bp

09-13-2013, 09:06 AM

Hi
I have been preparing libraries from approx 400ng total RNA, mRNA enrichment, using the ScriptSeqV2 kit.
At the end I amplify with 12 cycles and I get a profile with a 2nd peak around 550bp. I have not been able to get an explanation for this yet.

If somebody has an idea I would be grateful.

Thanks Ferdinand

P.S. Profile attached.
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Tags: problem, scriptseq
Olaf Blue

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Join Date: Nov 2010

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#2

09-13-2013, 10:25 AM

Did you quantify the input RNA for the Scriptseq library? The kit works best with 500 pg to 50 ng input.
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FerdinandvM

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Join Date: Jul 2013

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#3

09-14-2013, 05:32 AM

No, but assuming that mRNA will never be more than 10% of the total RNA I should be in range.
Could it be an issue with the fragmentation?
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ScriptSeq - 2nd peak @ 550bp

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