Path: utzoo!utgpu!news-server.csri.toronto.edu!bonnie.concordia.ca!thunder.mcrcim.mcgill.edu!snorkelwacker.mit.edu!bionet!CCIT.ARIZONA.EDU!P9323611 From: P9323611@CCIT.ARIZONA.EDU Newsgroups: bionet.molbio.genbank Subject: message Message-ID: Date: 19 Feb 91 00:02:00 GMT Sender: daemon@genbank.bio.net Lines: 46 Curious, I haven't been able to get a clear answer to a question I have had for almost a year now. I'm curious to know why researchers et. al. haven,t developed an in Vitro transcription system that could be utilized somewhat similar to the way the Polymerase Chain Reaction (PCR) is used. Instead of the crude extracts that are currently available on the market, can't the RNA polymerase from cells be purified and then used in a test tube containing: template DNA (containing the proper promoter sequence) RNA nucleotides ATP necessary ions this way a standardized method would be available -- in pre-prepared test tubes- for scientists to test specific transcription factors that are required for transcription of their particular gene. I know that different cells regulate their genes differently and by different mechanisms, but it seems to me that the basic necessary components of transcription should be available in a test tube. It seems like a standard system like this would supplement the current in vivo systems and would be relatively cheap. What is the limiting factor that is holding this technology back. Is it the purification of the RNA polymerase? Any information to these questions would be appreciated, John Pierpont, medical student University of Arizona