Sharp peak in amplified RNA library

gman85cini

Junior Member

Join Date: Sep 2014

Posts: 2
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#1

Sharp peak in amplified RNA library

09-23-2014, 07:03 AM

Hey everyone,

So i've been trying to make libraries from low-ish input (i.e.: 25-50ng total RNA) using Linearized DNA Amplification and NEB Next Ultra. RNA was polyA selected and fragmented. I'm getting this weird sharp peak at ~330 bp (see attached) after amplification (19 cycles) and I'm not quite sure what to make of it. Looking for any thoughts or suggestions. If you need more details let me know.

Thanks!

Giacomo
Attached Files

19 cycle post amp.pdf (787.3 KB, 199 views)
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nucacidhunter

Jafar Jabbari

Join Date: Jan 2013

Posts: 1250
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#2

09-23-2014, 06:46 PM

Providing more info would enable others to offer more comprehensive help. Not knowing the details of your library prep method, one possibility is contamination of libraries (or reagents used) with an amplicon possibly one flanked with Illumina adapters that has been amplified efficiently.
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Sharp peak in amplified RNA library

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