SEQanswers

Go Back   SEQanswers > General



Similar Threads
Thread Thread Starter Forum Replies Last Post
double peak in mRNA truseq kit ssing Sample Prep / Library Generation 17 03-16-2019 01:00 AM
mRNA from Bacterial sample for trueseq Garyron Sample Prep / Library Generation 9 08-16-2011 12:42 AM
Informatics cause of intra-sample variability, Illumina TruSeq mRNA eab Bioinformatics 0 07-13-2011 08:16 AM
Intra-sample variability, Illumina TruSeq mRNA eab General 0 07-12-2011 07:50 AM
questions about Illumina mRNA sample prep ik76 Illumina/Solexa 2 01-19-2010 03:38 AM

Reply
 
Thread Tools
Old 07-16-2011, 12:32 PM   #21
pmiguel
Senior Member
 
Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
Posts: 2,313
Default

Hi Eli,
In principle it should be possible to skip amplification altogether if you do qPCR on the unamplified library.
We got post-amplification yields in some cases that were less than pre-amplification. (These were DNA libraries.) Probably an issue with the ampure reaction clean-up. The libraries went on to produce plenty of sequence and none of them looked "bottomed out".
This is all just stuff to be aware of, in my opinion. The Illumina protocol should be fine in many cases. Might even be worth using their spike in controls. I personally an not yet able to get over the horror of adding extraneous DNA during library construction. But that is just me...

--
Phillip
pmiguel 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 05:53 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2019, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO