SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
16s updated NextSeq500 library prep for low concentration ok for MiSeq ? BioGenomics Sample Prep / Library Generation 4 11-17-2014 03:38 AM
Wang, 'Low Cost Library Construction...': Low Concentration MAdkisson RNA Sequencing 2 02-19-2014 02:33 PM
low library concentration biochembug Illumina/Solexa 0 08-02-2012 06:13 AM
Low concentration GA library Melanie Sample Prep / Library Generation 4 08-04-2010 01:34 PM
Anyone use low concentration for ChIP-seq prep? Audry888 Sample Prep / Library Generation 4 05-28-2010 11:37 AM

Reply
 
Thread Tools
Old 03-19-2015, 02:36 PM   #1
Normal
Junior Member
 
Location: USA

Join Date: Mar 2015
Posts: 1
Default Library prep/low concentration

Hi all,

I'm a new member in this forum. I'm having trouble with preparation of my sample using Nextera XT kit. Most of my samples came out with too low concentration after PCR cleaningup. I checked my starting DNA conc (e.g for 2.25ng/ul, I used 0.44ul sample). I don't have problem with my cleanup process but the library yield is always coming out too little for sequencing. Please, I'd appreciate anyone with valuable suggestions.
Thanks
Normal is offline   Reply With Quote
Old 03-19-2015, 03:12 PM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,238
Default

Two cause to consider:
1- Accuracy of quantification and method used
2- Library fragments were too short or bead ratio was low and fragments were removed during clean-up. Checking size distribution before clean-up should rule in or out this assumption.

For better help please post Electropherogram of few libraries.
nucacidhunter 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 03:40 PM.


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