I isolated RNA from NHBE cell line using qiagen minikit. The perform cDNA synthesis (1000 ng) and run qPCR using SYBR green with GAPDH primer (200 nM).
That is just background noise indicating that you have 0 amplification. Take a look at the scale on the Y-axis. If your positive control had any amplification, then the "noise" would be flattened and you'd see a curve.
Yeah, as Katie said, this is just background noise. Does your positive control (housekeeping gene) also look the same? if you didn't use that, try running again with positive control. Also, make sure your primer efficiency 99 or 100%
As the other guys Katie A S Burnette & Thanduanlung Kamei have stated, and make sure you have selected the right target (FAM) for your amplification since your melting curve is good, these might be readings from a wrong target also. & remember your amplification plot is not dependent on your melting curve.
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