SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
RNA-Seq: HITS-CLIP: panoramic views of protein-RNA regulation in living cells. Newsbot! Literature Watch 1 09-11-2015 12:48 AM
RNA-Seq: RNA-sequence analysis of human B-cells. Newsbot! Literature Watch 0 05-04-2011 03:50 AM
RNA-Seq: RNA-Seq Analysis of Sulfur-Deprived Chlamydomonas Cells Reveals Aspects of A Newsbot! Literature Watch 0 07-01-2010 03:40 AM
RNA-Seq: Transcriptome and targetome analysis in MIR155 expressing cells using RNA-se Newsbot! Literature Watch 0 06-30-2010 03:00 AM
RNA seq libraries from few hundred cells artem.men Sample Prep / Library Generation 2 03-02-2010 05:04 PM

Reply
 
Thread Tools
Old 12-27-2016, 06:46 PM   #241
Timer123
Junior Member
 
Location: Wuhan.China

Join Date: Dec 2016
Posts: 2
Default Tn5 assemble and single cell sequence

I am a student from Huazhong Agricultural University,Wuhan,China.Now I am working with the single cell RNA-seq library preparation,because we have so many sample to operate, we could not afford this by use standard Nextera kit,so we have to produce in-house Tn5 transposase by reference the article of "Tn5 transposase and tagmentation procedures for massively scaled sequencing projects"Picelli et al.Genome Research.2014.
In the assemble protocol,where I can get a most suitable the phosphorylation primer:
Tn5MErev, 5'-[phos]CTGTCTCTTATACACATCT-3',and is there any detail protocol or notes about Tn5 transposase assemble and purification ?
And for the smart-seq2, how to order the TSO primer also a problem in China, I know the Exiqon have the patent for the LNA,and order is very difficult. So how to order it faster or is there any good alternative company I can get this?
Thank you for your generous help!
Timer123 is offline   Reply With Quote
Old 12-30-2016, 02:06 AM   #242
Simone78
Senior Member
 
Location: Sweden

Join Date: Oct 2010
Posts: 173
Default

Quote:
Originally Posted by Timer123 View Post
I am a student from Huazhong Agricultural University,Wuhan,China.Now I am working with the single cell RNA-seq library preparation,because we have so many sample to operate, we could not afford this by use standard Nextera kit,so we have to produce in-house Tn5 transposase by reference the article of "Tn5 transposase and tagmentation procedures for massively scaled sequencing projects"Picelli et al.Genome Research.2014.
In the assemble protocol,where I can get a most suitable the phosphorylation primer:
Tn5MErev, 5'-[phos]CTGTCTCTTATACACATCT-3',and is there any detail protocol or notes about Tn5 transposase assemble and purification ?
And for the smart-seq2, how to order the TSO primer also a problem in China, I know the Exiqon have the patent for the LNA,and order is very difficult. So how to order it faster or is there any good alternative company I can get this?
Thank you for your generous help!
I replied you in a private message, take a look there!
Simone78 is offline   Reply With Quote
Old 01-02-2017, 12:03 AM   #243
Timer123
Junior Member
 
Location: Wuhan.China

Join Date: Dec 2016
Posts: 2
Default Tn5 assemble and single cell sequence

I have read that,it's very useful for me, thank you very much!
Timer123 is offline   Reply With Quote
Old 01-18-2017, 11:45 PM   #244
rnaseqmonkey
Junior Member
 
Location: Germany

Join Date: Feb 2014
Posts: 2
Default

Did anyone solve the mystery of bacterial rRNA contaminations yet?
We're also seeing this pop up inversely correlated to the endogenous RNA amount. We use Maxima H- RT for our libraries, too!
rnaseqmonkey 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 10:53 AM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2017, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO