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Periodic baseline fluctuation in melting curve


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#1 roryk

roryk

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Posted 05 August 2009 - 07:13 AM

Hi! First post on the forum.

I read through all of the topics on this board and did not see any that addressed a problem I am having. I am using SYBR green with ROX as the passive dye on the Bio-Rad DNA Engine Opticon 2 cycler. When I look at my melting curves, I have periodic fluctuations in the baseline from 60-90C, even in wells with no template. An example of the melting curve is here:

www.ice-nine.org/rory/qpcr_troubles.pdf

Any ideas about what I am doing wrong? This is not limited to just this well, it is all of the wells. I suspect that I did something like not have enough of a wait time between steps in the melting curve analysis or something like that. Does anybody using similar dyes with a similar machine have any recommendations about a proper curve to use?

Thanks!




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