SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
Staff Scientist Bioo Scientific Bioo Scientific Industry Jobs! 0 03-21-2013 12:39 PM
Bioo Scientific kits Turnerac0987 Sample Prep / Library Generation 22 11-15-2012 01:08 AM
Barcode ChIP-seq library using Bioo Scientific Kit inankai Sample Prep / Library Generation 5 11-17-2011 08:11 PM
Library prep from single-stranded DNA? genlyai Sample Prep / Library Generation 3 11-12-2010 12:26 PM
Bioo Scientific Chooses Eureka Genomics as its Next-Generation Sequencing Provider Bioo Scientific Vendor Forum 0 08-17-2010 06:48 AM

Reply
 
Thread Tools
Old 04-08-2015, 12:28 PM   #1
teofil
Junior Member
 
Location: Arkansas

Join Date: Mar 2015
Posts: 1
Default BIOO Scientific stranded RNAseq library prep

Dear members,

We are using the BIOO Scientific NextFlex rapid directional RNAseq kit to prepare RNAseq libraries and getting some unexpected results. Namely, in about half of our preps, TapeStation electropherograms show a marked peak at the low end of the library followed by a ~normal looking profile toward the higher end of the library (see attached pdf). In addition, and compared to nondirectional TruSeq preps routinely performed in the lab, we are getting quite low library yields (commonly ~ 1ng/uL and sometimes below that). The input RNA is 200 ng with RIN > 7. The RIN numbers are not great due to peaks from organelle rRNA (we think primarily chloroplast).

We suspect that we might be doing something wrong, but we are unsure what. Any suggestions would be great (fragmentation, PCR, Ampure cleanups).

Also, has anyone had experience sequencing libraries looking like this? We're planning to sequence on a HiSeq 2000.

Thanks!
Attached Files
File Type: pdf R134_library.pdf (214.0 KB, 27 views)
teofil is offline   Reply With Quote
Old 04-09-2015, 10:48 AM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,231
Default

I wonder if you have asked Bioo techsupport for troubleshooting. Technically those libraries can be sequenced, but they are less likely to provide publication quality data.
nucacidhunter is offline   Reply With Quote
Reply

Tags
nextflex, rnaseq, stranded library prep

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 11:01 PM.


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