HELP for PCR!!!

Pete pkursula at
Fri May 30 02:49:00 EST 1997

I suppose that the large band is your plasmid, not an amplification
product and the fast one might be dNTPs/oligonucleotide primers. Thus, it
may be that there is something that interferes with the specific
amplification of your target, or amplification in general.

Just a guess, 

On 29 May 1997, ma wrote:

> Dear Netters:
> I met a problem recently in PCR a plasmid. I had got the correct product (about 800bp) 
> using the same plasmis, Taq enzyme, primers and other PCR components before. But now I 
> only get a large and wead band (more than 3kb), in the same time I found  a pecuilar 
> phenomenon----in gel analysis, there is a band (I call it, but it is not the real DNA 
> band) runing faster than bromopheonol blue dye, yet it can not be seen in normal light, 
> we can only see "this band" under UV light yet it is look very like bromopheonol blue 
> dye. Do any PCR experts have ideas about this, What is "this band" represent? and how to 
> get rid of it? Since the funding is tight in our lab, I do not like to make a serial of 
> test to investigate it, so your experiences will be very valuable. Thanks very much.
> Ma

More information about the Methods mailing list