SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
TruSeq mRNA Elution Buffer eyoungman Sample Prep / Library Generation 0 04-06-2017 06:54 AM
Illumina TruSeq resuspension Buffer lvandiepen Illumina/Solexa 6 01-03-2017 04:14 PM
Ampure XP DNA rebinding if accidentaly added RSB Ancestro Sample Prep / Library Generation 1 11-06-2015 06:19 PM
Running BioA on RNA/cDNA samples in lysis/RT buffer SunPenguin Sample Prep / Library Generation 0 09-21-2015 12:42 PM
End repair of TruSeq RNA and TruSeq mRNA stranded ykgenome Sample Prep / Library Generation 2 05-15-2014 12:01 AM

Reply
 
Thread Tools
Old 11-13-2018, 06:58 AM   #1
GenomeM
Junior Member
 
Location: MDE

Join Date: Nov 2018
Posts: 3
Default Resuspension Buffer (RSB) added to RNA samples in Truseq mRNA Protocol

Hello ,

can Resuspension buffer prevent mRNA from binding to RNA beads in first step of Truseq protocols?
it was used instead of water to bring volumes of samples to 50uL

if any one can provide the contents of the buffer too ?

thanks alot in advance,

GM
GenomeM is offline   Reply With Quote
Old 11-13-2018, 07:26 AM   #2
GSviral
Member
 
Location: UK

Join Date: Dec 2014
Posts: 31
Default

Hi,

I'm not too sure about the resuspension buffer preventing the binding to the RNA beads, but in any TruSeq protocol I've used the buffer was used to remove the libraries from the beads after binding and washing so it could possibly inhibit binding but someone else will have to confirm that.

In a previous position we had a homemade RSB recipe which was just TE buffer + Tween-20 (I can't remember off the top of my head but it might have just been 1% Tween-20) and it worked fine. I don't know the actual chemical make-up though, sorry.
GSviral is offline   Reply With Quote
Old 11-13-2018, 08:22 AM   #3
GenomeM
Junior Member
 
Location: MDE

Join Date: Nov 2018
Posts: 3
Default

GSviral

Thank you for your reply
Yes it has multiple use to release templates off the beads after thermalcycling episodes
So that is what hit me to ask is it by composition unbind tue teplates or the heat is the starter

Thanks alot again

GM
GenomeM is offline   Reply With Quote
Old 11-16-2018, 02:18 AM   #4
GenomeM
Junior Member
 
Location: MDE

Join Date: Nov 2018
Posts: 3
Default

Quote:
Originally Posted by GSviral View Post
Hi,

I'm not too sure about the resuspension buffer preventing the binding to the RNA beads, but in any TruSeq protocol I've used the buffer was used to remove the libraries from the beads after binding and washing so it could possibly inhibit binding but someone else will have to confirm that.

In a previous position we had a homemade RSB recipe which was just TE buffer + Tween-20 (I can't remember off the top of my head but it might have just been 1% Tween-20) and it worked fine. I don't know the actual chemical make-up though, sorry.
GSViral

thank you for sharing your notes, I have completed the prep the RSB did not affect the binding specially after thermal cycling step which might enhance the binding of the poly-A to the T-tail of the beads.

GM
GenomeM is offline   Reply With Quote
Old 11-19-2018, 01:19 AM   #5
GSviral
Member
 
Location: UK

Join Date: Dec 2014
Posts: 31
Default

Hi GenomeM,

Thanks for giving an update on this, it's good to know that the RSB didn't affect the binding! I'll keep that in mind for the future.

Cheers.
GSviral 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 01:19 AM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2018, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO