Silanized equipment when purifying tRNA? - (Oct/23/2012 )
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
-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.
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.