Methods for fluorescence data correction

Abstract

Method of processing real-time PCR data, comprising: c) receiving a plurality of fluorescence melting curve data of real time PCR-experiments performed by a real-time PCR device with at least two fluorescence channels, and configured to perform the following steps multiple times, while increasing a temperature: i) at first moments in time measuring a first temperature value and a first radiation value corresponding to a first fluorescence channel; ii) at second moments in time measuring a second temperature value and a second radiation value corresponding to a second fluorescence channel; d) storing the plurality of temperature values and radiation values; e) determining a plurality of time-shifted second radiation values by linearly interpolating between two measured second radiation values, using weighting factors defined by the measured temperature values; f) after performing step e), calculating color corrected first radiation values, and color corrected second radiation values.