Go Back   SEQanswers > Literature Watch

Similar Threads
Thread Thread Starter Forum Replies Last Post
home made EZ-Tn5-Nextera adapter complex? silin284 Sample Prep / Library Generation 1 10-01-2019 11:02 AM
Dealing with ">" and "<" signs in mapping column in RNA-Seq pileup file for RNA-edit slp Bioinformatics 0 03-21-2014 10:18 AM
Home-made ABi SOLiD Total RNA-Seq ?? dcyounge RNA Sequencing 5 04-03-2012 05:39 AM
normalizing RNA-seq data to "unique transcript length" instead of "transcript length" lmc Bioinformatics 2 06-23-2010 11:45 AM
First Helicos Publication! Single Molecule DNA Seq of a "Viral" Genome ECO Helicos / Direct Genomics 15 09-05-2008 07:05 AM

Thread Tools
Old 07-31-2014, 08:49 AM   #1
Senior Member
Location: Basel (Switzerland)

Join Date: Oct 2010
Posts: 208
Default "home-made" Tn5 transposase for single-cell RNA-seq

here is our paper on Tn5 transposase production which doesn´t require the purchase of an expensive Nextera kit from Illumina:

Massively parallel DNA sequencing of thousands of samples in a single machine-run is now possible, but the preparation of the individual sequencing libraries is expensive and time-consuming. Tagmentation-based library construction, using the Tn5 transposase, is efficient for generating sequencing libraries but currently relies on undisclosed reagents, which severely limits development of novel applications and the execution of large scale projects. Here, we present simple and robust procedures for Tn5 transposase production and optimized reaction conditions for tagmentation-based sequencing library construction. We further show how molecular crowding agents both modulate library lengths and enable efficient tagmentation from sub-picogram amounts of cDNA. Comparison of single-cell RNA-sequencing libraries generated using produced and commercial Tn5 demonstrated equal performances in terms of gene detection and library characteristics. Finally, as naked Tn5 can be annealed to any oligonucleotide of choice, for example molecular barcodes in single-cell assays or methylated oligonucleotides for bisulfite sequencing, custom Tn5 production and tagmentation enables innovation in sequencing-based applications.
Simone78 is offline   Reply With Quote
Old 08-05-2014, 11:37 PM   #2
Registered Vendor
Location: Eugene, OR

Join Date: May 2013
Posts: 523

Nice work! Lots of great details beyond the DIY aspect.
Providing nextRAD genotyping and PacBio sequencing services.
SNPsaurus is offline   Reply With Quote
Old 08-21-2014, 02:38 PM   #3
Location: Cambridge

Join Date: Nov 2012
Posts: 10

Nice work Simone!

I would just have one question why did you decide to have your TN5 wild type with respect to the M56 mutation?

Luka is offline   Reply With Quote

nextera sample prep, rna-seq, single cell, tn5

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 03:51 AM.

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