I am having a small problem with my SYBR green qrtpcr. I have 4 primer sets running against 2 different cDNA samples. The primers are all at the same concentration (300 nm). I prepared both cDNA samples side by side, in exactly the same way and loaded 10ng of each.
The problem is, when I look at the melt curve, I'm seeing a decrease in Tm of the product of about 1 degree C from one cDNA to another. This Tm shift occurs for all 4 primer sets. In both cases, there is a single sharp peak, so I'm doubting I have primer dimers. Does anyone have any ideas? I've used these primers before on different samples, and the melting curves always exactly coincided, no matter what the cDNA was. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
