The algorithms implemented in SoFAR (distributed by Metralabs) allow fully automatic analysis of real-time PCR data obtained with a Roche LightCycler® (Roche Diagnostics) instrument. The software yields results with considerably increased precision and accuracy of real-time quantification. This is achieved mainly by the correction of amplification independent fluorescence signal trends and a robust fit of the exponential phase of the signal curves. The melting curve data are corrected for signal changes not due to the melting process and are smoothed by fitting cubic splines. Therefore, sensitivity, resolution, and accuracy of melting curve analyses are improved (Wilhelm et al., 2003).
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