More RT PCR questions: - can I amplify "everything?" (Nov/01/2005 )
I am extracting RNA from some tricky stuff and want to use RT PCR to amplify any RNA that might be present. Is this a possibility? I feel comfortable converting the RNA into cDNA, but not with the amplification. It seems that the specificity of the primers would limit what gets amplified. Can I just use random primers again? Or should I pick a housekeeping gene to look at? B-actin?
Also, which kits do people prefer for the reverse transcription (M-MLV, sensiscript, first strand)?
Thanks!!
-jwahini-
QUOTE (jwahini @ Nov 1 2005, 12:16 PM)
I am extracting RNA from some tricky stuff and want to use RT PCR to amplify any RNA that might be present. Is this a possibility? I feel comfortable converting the RNA into cDNA, but not with the amplification. It seems that the specificity of the primers would limit what gets amplified. Can I just use random primers again? Or should I pick a housekeeping gene to look at? B-actin?
Also, which kits do people prefer for the reverse transcription (M-MLV, sensiscript, first strand)?
Thanks!!
Also, which kits do people prefer for the reverse transcription (M-MLV, sensiscript, first strand)?
Thanks!!
Hi. My lab has been using Applied Biosystems' high capacity cDNA archive kit and/or the reverse tanscription kit. They work very well for us.
-kiwi-
I will take amikins suggestion from another post and recommend that you make a cDNA library...
-beccaf22-