SEQanswers

Go Back   SEQanswers > Applications Forums > Sample Prep / Library Generation



Similar Threads
Thread Thread Starter Forum Replies Last Post
Help with TruSeq gDNA library prep BTS Illumina/Solexa 45 02-22-2017 10:09 AM
A really bad library? FrSr RNA Sequencing 2 02-11-2011 01:45 AM
bad library prep results dina Sample Prep / Library Generation 3 09-16-2010 05:00 AM
SPRI-TE Illumina library preps GW_OK Sample Prep / Library Generation 5 08-23-2010 09:13 AM
Ligase for DIY GAII library preps nickb Sample Prep / Library Generation 4 03-05-2010 10:36 AM

Reply
 
Thread Tools
Old 11-04-2011, 02:06 AM   #1
lukas1848
Member
 
Location: Germany

Join Date: Jun 2011
Posts: 54
Question Why and how are gDNA contaminations in RNAseq library preps bad?

Hi,

the title says it all.

I am doing Trizol extractions of total RNA for subsequent RNAseq library preps from small tissue samples. I'm wondering if DNAse treatment and/or LiCl precipitation are necessary to remove residual gDNA from my total RNA extracts.

Once again I'm referring to the Sengupta et al paper (in which they describe the preparation of RNAseq libraries from 10 ng total RNA) as their initial RNA purification is remarkably simple.
lukas1848 is offline   Reply With Quote
Old 11-04-2011, 03:51 AM   #2
protist
Senior Member
 
Location: Ireland

Join Date: Jan 2009
Posts: 101
Default

Hi,
Contaminating gDNA can be carried through the RNAseq library generation. For Eukaryotic libraries it can manifest as increased intergenic or intron reads that are not representative of the actual transcriptome. If you are concerned my advice would be check your RNA with a quick RT-PCR.
protist is offline   Reply With Quote
Reply

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off




All times are GMT -8. The time now is 06:20 AM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2020, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO