Hi all,
I'm having problems with my beta actin gene melting curve. In the past, the melting curves were working well (Fig. 1). Recently, I've faced with some problems (Fig2).
1- The y axis (fluorescence) shows different levels, even comparing the same sample (different technical replicates) - Pipettes are calibrated.
2 - The beginning of the curve are messy and high, comparing with my first figure. The primers are new.
Anyone have any idea what is the problem and can help me?
Thank you so much