From owner-drosophila@net.bio.net Mon Mar 27 23:00:00 1995
Path: biosci!galaxy.ucr.edu!ihnp4.ucsd.edu!sdcc12!jeeves!dma
From: dma@jeeves.ucsd.edu (Danuta Maria Antosiewicz)
Newsgroups: bionet.drosophila
Subject: pcr problem with genomic fly
Message-ID: <82084@sdcc12.ucsd.edu>
Date: 28 Mar 95 00:57:27 GMT
Sender: news@sdcc12.ucsd.edu
Organization: University of California, San Diego
Lines: 5
Nntp-Posting-Host: jeeves.ucsd.edu

To anyone on the net who knows:
	
		I have been attempting to PCR a fragment on the Drosophila Simulans TRA gene with almost no luck.  My primers are both 17 bases long and they span a segment of about 800 bases.  I've tried many different annealing temperatures from 53 - 65 degrees Celsius, and have also tried different concentrations of genomic DNA,  I've run 25 cycles, 30 cycles, and 35 cycles, and I still can't get the damn thing to work.  Does anyone know anything I don't know about genomic fly DNA and the effective PCR of these DNA? 

 Please respond.  My email address is:  jalin@ucsd.edu    Thanks.

.