SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
TruSeq or Nextera kit for library prep? rooty Sample Prep / Library Generation 12 10-29-2013 06:02 AM
Nextera vs Illumina Library Prep maggie Illumina/Solexa 6 03-01-2012 06:01 PM
Illumina Trueseq RNA prep kit problems cburke11 Sample Prep / Library Generation 19 09-27-2011 08:42 AM
Nextera vs Illumina Library Prep maggie Illumina/Solexa 2 12-30-2010 09:58 AM
Homemade illumina oligos and library prep kit elly Illumina/Solexa 3 03-24-2009 08:05 AM

Reply
 
Thread Tools
Old 10-24-2011, 01:34 PM   #1
fgoetz
Junior Member
 
Location: Providence, Rhode Island

Join Date: Jan 2011
Posts: 1
Default cDNA prep for Illumina using the Nextera kit

Hello!

We are looking to streamline our cDNA sample prep for the HiSeq and many of our samples have very low quantities of RNA and are very hard to collect. So far we have tried the Nugen Ovation V2 kit for cDNA synthesis and finished with the TruSeq kit. We are waiting to see what the data look like but the yields look great!

Has anyone tried working up cDNA with Nextera? If so, how did it go and do you have any advice on this?

Has anyone tried following the Nugen Ovation V2 with the Nextera kit? If so, I would appreciate hearing any experiences, advice or suggestions.

Thank you for your time!
fgoetz is offline   Reply With Quote
Old 03-16-2012, 12:06 PM   #2
kmkocot
Member
 
Location: Alabama

Join Date: Jun 2009
Posts: 48
Default

I have the same question. I'm working with microscopic animals and making cDNA libraries using the Clontech SMARTer kit but it just doesn't yield enough ds cDNA for TrueSeq library prep.

I imagine that the Nextera kit would result in bias towards sequencing the middle regions of transcripts? Illumina's FAQs say that Nextera yields fragments between 300 and 1000 bp so I'm worried that some short transcripts might not get tagmented at all.
kmkocot is offline   Reply With Quote
Old 03-16-2012, 12:24 PM   #3
kmkocot
Member
 
Location: Alabama

Join Date: Jun 2009
Posts: 48
Default

After doing a little more searching it looks like it can be done!

http://epicentral.blogspot.com/2010_08_01_archive.html

The Myers lab at the Hudson Alpha tested it and got good sequencing results with human mRNA.

Thanks,
Kevin
kmkocot is offline   Reply With Quote
Old 07-09-2012, 06:34 PM   #4
kmkocot
Member
 
Location: Alabama

Join Date: Jun 2009
Posts: 48
Default

Just a bump...

Has anyone tried the new Nextera XT sample prep kit with cDNA? They say it's good for small genomes, plasmids, and amplicons but nothing is said about cDNA.

Thanks,
Kevin
kmkocot is offline   Reply With Quote
Reply

Tags
cdna, nextera, nugen, ovation, truseq

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 10:03 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2018, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO