SEQanswers

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



Reply
 
Thread Tools
Old 05-04-2015, 11:29 AM   #1
clintp
Member
 
Location: Georgia

Join Date: Apr 2013
Posts: 17
Default Best plasticware?

What are people using as best practice for plasticware in NGS library preparation? For 1.5 and 0.6 ml tubes, I've been using Eppenforf's LoBind tubes (library storage and dilution). I primarily use strip tubes for library generation, but I haven't found any 0.2mL tubes that are meant to prevent DNA sticking. I wonder if anybody has any input on what plastic is best for low-concentration RNA/DNA, and if anyone has any specific evidence to support it. Thanks!
clintp is offline   Reply With Quote
Old 05-08-2015, 12:52 AM   #2
Simone78
Senior Member
 
Location: Bonn (Germany)

Join Date: Oct 2010
Posts: 186
Default

Hi,
I had the same problem and I searched for a long time some low-binding 0.2 ml tubes or plates without success. Then I found a very old paper (attached) where they showed that, when adding some detergent to the solutions/mixes, the adsorption of DNA to the tube was negligible. We are doing single-cell RNA-seq and we have 0.1% Triton in the lysis buffer. We manage to achieve a better sensitivity than the commercial kits, therefore I believe that having some detergent is enough for our purpose and we donīt need low-binding tubes.
I hope that this helps!
Best,
Simone

Last edited by Simone78; 08-24-2017 at 05:42 AM.
Simone78 is offline   Reply With Quote
Old 09-14-2017, 02:03 PM   #3
COcard
Junior Member
 
Location: denver

Join Date: Jan 2013
Posts: 1
Default

Hey there, I know this is an old thread, but what is the citation for the "very old paper" you mention in your reply to clint ?
COcard is offline   Reply With Quote
Old 09-15-2017, 12:10 AM   #4
Simone78
Senior Member
 
Location: Bonn (Germany)

Join Date: Oct 2010
Posts: 186
Default

Quote:
Originally Posted by COcard View Post
Hey there, I know this is an old thread, but what is the citation for the "very old paper" you mention in your reply to clint ?
Please find it attached.
Best,
Simone
Attached Files
File Type: pdf detergent prevents adsorption of DNA to tubes.pdf (191.3 KB, 15 views)
Simone78 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 02:59 PM.


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