PCR problem

Jun Zhang junz at sfu.ca
Mon Mar 23 12:12:35 EST 1998


> 
> 
> 	I have been trying to PCR a 2kb fragment from a phage clone I am
> working with. I have done this succesfully in the past and amplified a
> single band using a 2 min extension and 55 celsius annealing temp.
> However now that I want to repeat this on the same template I get the
> correct size band but with a smear and several faint bands. This smear
> is across the size range of 2kb and above. Why am I suddenly getting
> this smear when all the parameters are the same as before? 
> 
> Thanks Richard
> 
> 
> 
hi, Richard:

i think there is too much template. you can try to dilute your template.
and don't use too much enzyme.

good luck.

mark zhang
biology dept.
simon fraser university
burnaby bc v5a 1s6
junz at sfu.ca




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