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RT-PCR contamination in negative control.

PCR contamination negative control

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#1 Smitha7

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Posted 28 March 2012 - 10:42 AM

So I have been working on these RT-PCR experiments for a couple of months now and with each experiment except for one I am getting a band that is the same as my product size in my negative control which is the -RT control. I take all procautions necessary that I can do given our limited acess to fume hoods. So I do all my PCR experiments in a seperate place in the lab. I have changed all my primers, DNTPS, water and yet I still get contamination in some of my samples but not in all my samples. I do use the tubes that are connected but I also have acess to tubes that aren't connected. I really don't know how I am getting contamination in these controls? We don't have access to filter tips in the lab of UV light. So I am somewhat pretty limited in controling some aspects.

#2 Trof

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Posted 28 March 2012 - 02:17 PM

Your negative control is RT- control? Do you also have NTC (non-template control) having the same band?
RT- can be contaminated with DNA giving a positive signal, if your primers are within one exon.

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