missing bands and ghost bands

Christian Praetorius via methods%40net.bio.net (by prae from gmx.net)
Tue Sep 30 04:09:19 EST 2008


"Manuel Gadin" <manuelg from telus.net> wrote:

First: Could you please set the length of your line to something like
70 characters? That makes it much easier to read.

>The method/procedure , master mix , gels and thermocycler that I am using is the same that my 
>co-lab workers are using. Having said that, my techniques could be the only culprit. I am the only 
>one that cannot get consistent results. I am failry new to PCR.Sometimes, I would be missing bands
>in my positive control and also I would get ghost bands. Few of my co workers had watched me run 
>the procedure and the only comment I got was maybe the parafilm degraded my DNA bands. The 
>parafilm would sometimes stick to my pipet tips when I am mixing the samples with the loading dye.
>Other than that, they cannot offer any solution.

I don't believe that Parafilm makes any trouble here. I am using the
same method for years, and when I had problems, it was always
something different, although it is not always easy to find.
Please describe your protocol in detail, what is the origin of your
DNA and so on. And if possible, show a photo of one of your gels.

Christian

-- 
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