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contamination in real-time pcr negative control


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#1 flashboy

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Posted 11 March 2005 - 02:10 AM

hi there,

i am noew constantly getting a late peak (30+ cycles) in the negative controls for my real-time pcr (SYBR Green, iCycler). i have even bought in new nuclease free water to run as my negative contro, and still get these peaks... does this mean my primers are contaminated????

#2 methylnick

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Posted 12 March 2005 - 08:48 PM

Your working primer stock could well be contaminated, when you say it's great than cycle 30, how much greater? if it's high 30's ie greater than 35 then I would not worry too much about it.

One other thing is to check the reaction by running it on a gel, if a distinct band comes up in your negative control(and it looks like the test lanes) well that would answer your question.

Good luck!

Nick :angry:




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