TaqMan Contamination

Robert Chadwick rchadwick at nce-mail.com
Tue Dec 12 13:05:02 EST 2000


If you are using the 5700, I bet you are doing TaqMan using SYBR green.  One
thing to watch out with SYBR Green is that it intercalculates between any
double stranded DNA molecule, so you will also detect artifacts such as
primer-dimers that form in the PCR.  Thus the contamination in your no
template controls is most likely primer-dimers amplifying.
The only way to prevent primer-dimers from forming is by proper design of
your PCR primers so that they won't have regions where they can bind
together.  Hot start PCR also helps to reduce primer-dimer formation but
this is standard practice in TaqMan and you are probably already using it.

>Subject: Re: TaqMan Contamination
>From: Rob Schweininger (RSchweininger at galileolabs.com 
>Date: Tue 12 Dec 2000 - 01:08:44 GMT 

>I have recently started using the 5700 Taqman and I am experiencing
problems 
>with amplification of the non template control. I have changed reagents, 
>changed prep space area, and cleaned the entire workspace environment to no

>avail. It seems that the only solution that I have come across via emails 
>is the use of the AmpErase system. I will post another message if the 
>AmpErase is successful. I am glad to know that I am not the only person 
>experiencing this problem. I was about to lose my mind! 


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