Resuspension Buffer (RSB) added to RNA samples in Truseq mRNA Protocol

[GenomeM]

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

[GSviral]

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.

[GenomeM]

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]

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

[GSviral]

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.