pcr prob

ChenHA via methods%40net.bio.net (by hzhen from freeuk.com)
Sun Feb 24 17:45:33 EST 2008


Aawara Chowdhury wrote:
> In <1203860754.14229.0 from proxy00.news.clara.net>,
>  ChenHA <hzhen from freeuk.com> wrote:
>
> The strong band is unlikely to be "just your primers".  As someone 
> earlier in this thread described, the strong band is what has 
> quixotically been labeled as "primer dimer" - the result of DNA synthesis 
> after a primer anneals (partially) to itself or the other primer in the PCR.
> 
> Loading 5 or 10 picomoles of each primer on an agarose gel by itself
> will not give a band that fluoresces brightly in the presence of 
> ethidium bromide.
> 

How do you know how much primers the original poster added?  It is for 
the original poster to comfirm or deny, or check by runnning the gel (as 
I stated, by using the same amount), not for you to assert.  Whoever 
gives you the idea the 5 or 10 picomoles is what everyone uses?  I 
don't, and I have done perhaps thousands of successful PCR reactions. 
And they do fluoresces, how brightly of course depends on how much you add.

As it happens, I see this kind of things quite often, from my own PCR.


> AC


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