baseline correction - (Sep/19/2005 )
I've been doing quite a bit of qPCR lately to detect gDNA contamination levels in my RNA preps. I'm using FAM labeled LUX primers on the MJ Research Opticon 2, and I'm a little confused about the baseline correction
Most of the time I don't need to do any baseline correction (usually just set to global minimum) but sometimes I get some samples where the fluorescence increases massively in the first cycle, but then increases again later on. When this happens the only way I can make sense of the results is to set the baseline correction to average over cycle range and set the range to all cycles, ie after the amplification has plateaued. The melting curves are all perfect
My question is, is there anything wrong with doing this or am I falsifying my results?
wow, thanks for the help guys