I made an RT prep from cultured cells and am running PCR on the resulting cdna. Each time I run Bact on the same samples using the same primers and conditions I get a different result. Most confusing of all is that my CT values are increasing every time I run the assay. Has anyone had any experiences like this and know what the problem may be?
Alli Mathiesen
I wouldn't expect degradation to result in steadily increasing CT values, and I made aliquots to limit freeze/thaw. In some cases, the successive runs were hours apart with the exact same reagents and my controls showed no variation between runs.
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