PCR Trouble

Scott Wildenberg DGM-IHG wilden at lenti.med.umn.edu
Fri Jun 3 13:09:57 EST 1994


I have been trying to amplify a region of human genomic DNA.  I have 
amplified it successfully in the past, but, under the same conditions, 
some new samples are not working.  I have done a phenol/CHCl3 extraction 
and ethanol precipitation to clean them up, run the reactions with MgCl2 
concentrations ranging from 0.5 mM to 4.0 mM, and tried increasing the 
amount of DNA.  I got a very faint band when I increased the amount of 
DNA to 400ng (50 ul rxn).  I figure it must be something about the 
samples because I have run positive controls, and they always look great.
Does anyone have any advice on what I can do?


Thanks,

Scott C. Wildenberg

Division of Genetics and Metabolism   	Voice (612) 625-5628
University of Minnesota               	Fax (612) 624-6645
Box 485 UMHC                            Email wilden at gene.med.umn.edu
Harvard Street at East River Road     
Minneapolis, MN  55455                   



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