Question: (Closed) RNA-seq library/Gel purification/bioanalyzer
0
gravatar for linjc.xmu
4.3 years ago by
linjc.xmu10
linjc.xmu10 wrote:

Hi all, I prepared RNA-seq library following SMART template switch protocol. This protocol worked well with my samples. But I don't know what happened these days. When I got cDNA, I purified it by AMPure beads. And then do PCR. I ran the PCR product on 2% agarose gel and do size selection from 300-500bp. I can see a good smear concentrated at 400bp. I use ZYMO kit to purify the target band. Then I checked the purified product on Bioanalyzer, the distribution was much broader than 300-500bp (almost 200-1000bp or higher). I also ran this purified product on 2% gel again, the distribution became broader as the Bioanalyzer showed. Have anybody met with this kind of problem?

Thank you very much.

rna-seq next-gen • 1.5k views
ADD COMMENTlink written 4.3 years ago by linjc.xmu10
1

Hello linjc.xmu!

We believe that this post does not fit the main topic of this site.

Not linked to bioinformatics

For this reason we have closed your question. This allows us to keep the site focused on the topics that the community can help with.

If you disagree please tell us why in a reply below, we'll be happy to talk about it.

Cheers!

ADD REPLYlink written 4.3 years ago by WouterDeCoster44k

You should ask this on seqanswers instead, the community there is better able to field library prep. questions.

ADD REPLYlink written 4.3 years ago by Devon Ryan97k
Please log in to add an answer.
The thread is closed. No new answers may be added.

Help
Access

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.
Powered by Biostar version 2.3.0
Traffic: 1481 users visited in the last hour