HELP for PCR!!!
pkursula at cc.oulu.fi
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.
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