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

Similar Threads
Thread Thread Starter Forum Replies Last Post
Adapting TruSeq RNA-Seq Protocol JayS Sample Prep / Library Generation 3 04-13-2011 06:58 PM
mRNA library prep alternative protocol lvandiepen RNA Sequencing 0 01-18-2011 04:19 PM
Illumina directional RNA-seq protocol Herve Illumina/Solexa 10 06-13-2010 07:18 AM
Illumina mRNA-seq protocol Xi Wang Sample Prep / Library Generation 0 01-02-2010 06:13 AM

Thread Tools
Old 02-25-2010, 07:42 AM   #1
Junior Member
Location: London, UK

Join Date: Jan 2010
Posts: 9
Default illumina alternative v1.5 protocol for small rna seq vs. the standard protocol

Hi All,
I was just wandering whether anybody had successfully used the illumina alternative v1.5 small RNA sample prep protocol . We tried but the only thing we could detect was adapter contamination.
If anybody has used it successfully can they comment on the 3' adapter ligation protocol ( the one using T4 RNA ligase 2 truncated). I was wandering why the portocol uses such a high Mg concentration ( 10mM I believe) and does not use PEG.
ik76 is offline   Reply With Quote
Old 03-25-2010, 02:24 PM   #2
Bioo Scientific
Registered Vendor
Location: Austin, Tx

Join Date: Oct 2009
Posts: 99
Default Troubleshooting v1.5 sample prep protocol

Where did you get your adapter? What concentration of adapter did you use and how many units of ligase did you use? Adding too much adapter may lead to primer dimers and other artificial PCR products.

We have used the v1.5 sample prep protocol using our own enzyme, AIR Ligase, a truncated T4 RNA Ligase 2 successfully with our adenylated adapters. We would be happy to send you our recommended protocol and a sample of T4 RNA Ligase 2 (truncated).

Dawn Obermoeller
Bioo Scientific

Last edited by Bioo Scientific; 09-15-2015 at 10:49 AM.
Bioo Scientific is offline   Reply With Quote

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 01:43 PM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2021, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO