QC-PCR problem

Bilinski Robert bilinskr at ERE.UMontreal.CA
Mon Apr 3 22:39:37 EST 1995


	Hello, i'm using a friend's account to write this, feel free to 
send him any and all replies (as well as posting them on the newsgroup 
for the benefit of others...). Oh... and i must warn you that my english 
is not the best in the world...

	I'm trying to develop a competitive PCR method to quantify cell
characterized by translocation (14;18). I cloned a PCR product of a cell
line harboring this translocation, and now I must purify it and dilute it
to use it like an internal control. And I had some bugs with my plasmid's
purification (I have ribonucleotides and my 260\280 D.O. are 1.7) I think
i know why it's difficult for me to obtain a fonctionnnal dilution. It's
probably due to missquantification of my plasmid DNA stock. 

	Are there any other possible explanations for this phenomena? 
Could someone suggest a possible solution to this problem?

	Other usefull info:
		- I used a QIAGEN kit for maxiprep
		- I isolated the digested plasmid from agarose gel to 
obtain purer DNA but I failed in this attempt. (this was the solution i 
tried when i saw a RNA and ribonucleotide contamination in a DNA stock 
solution)


	I would appreciate any help anyone can offer...


						Thanks in advance
						Brigitte

ps: i"m also looking for any ressources available for people doing PCR in 
the Montreal region (province of Quebec, Canada), or maybe the names of 
other researchers (or groups of) in the same field close by... I'm 
presently the only one in my research center involved in such work (HMR, 
affiliated with The Universite de Montreal)  



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