Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Silanized equipment when purifying tRNA?


  • Please log in to reply
1 reply to this topic

#1 AaronF

AaronF

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 23 October 2012 - 01:58 PM

Our lab works extensively with tRNA and our PI is amazed that people have quit silanizing the things that come into contact with the tRNA during the purification process. I am a new member to this lab, but from what I can tell people did not like working with the silanizing agents (dimethyldichlorosilane and methylene chloride, there is some Sigmacote around but it seems everyone silanized with the previously mentioned solvents) and have sinced moved away from doing so. I am wondering if I should start silanizing equipment for the purification of in-vitro transcribed tRNA, or if silanization less necessary with the recent and vast improvements of laboratory plastics. As it stands, our general protocol is:

-In-vitro transcription of tRNA
-Ethanol precipitation and resuspension of pellet
-Urea-PAGE of tRNA
-Extract band of tRNA
-Crush extracted gel with glass (non-silanized) rod
-Elu-Trap purification
-Ethanol precipitation
-Resuspend and fold tRNA

My question is, would my yields be greatly improved by silanizing the microcentrifuge tubes and glass rods used in this purification? Or would the increase in yield be negligible? I'd rather not have to deal with the solvents involved in silanizing if I don't have to, but if it increased yields enough to offset the cost in reagents I could overcome my dislike of silanizing these things.

#2 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,504 posts
252
Excellent

Posted 23 October 2012 - 03:49 PM

You can purchase silanized tubes, which may be a better approach if you need only a few. I don't know about silanized glass rods, however.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.