RNA probe purification - (Feb/07/2001 )
hi,
Is there anyway to purify an RNA probe after in vitro transcription without using tRNA as a carrier for precipitation.
vimala
Try using one of the many spin columns found on the market. I've found pharmacia's s-200 columns to work well.
you can also purify you RNA transcripts using PAGE gel, cut the band and elute over night at 37C in elute buffer and use it without further treatment. Recently I used this method to prepare my RNA probe for RNase Protection analysis and it works quite well.
Good luck
| QUOTE ( @ Feb 8 2001, 11:00 PM) |
| you can also purify you RNA transcripts using PAGE gel, cut the band and elute over night at 37C in elute buffer and use it without further treatment. Recently I used this method to prepare my RNA probe for RNase Protection analysis and it works quite well.<p>Good luck [/quote] Hi, I was just wondering if you could make available your protocol for band extraction? Thanks Noel -Noel- I work with almost nothing but RNA and it is pretty easy. UV shadow or radio label your RNA cut the band out and either place it in TE and 0.1 %SDS overnight or electroelute it (can be done in dialaysis tubing and a agrose gel box). If you electroelute you will need to concentrate (butanol or microcon). If you need it to be super clean (NMR etc) then use a A25 DEAE (sigma). -5900P1- |