A method for quantitation of biological material in a biological sample is provided. The method includes receiving amplification data from amplification of a first and a second reference sample and receiving amplification data from amplification of a biological sample. The method further includes determining an efficiency from the received amplification data from amplification of the first and second reference sample. The method includes determining a relative PCR efficiency for the biological sample. Next, the method includes determining a quantity of biological material in the biological sample using the relative PCR efficiency.