Methods and systems for identifying the quantitation cycle for a PCR amplification reaction

摘要

Methods and systems for identifying a quantitation cycle (Cq) for a PCR reaction that includes fitting a line having a plurality of line segments to data points associated with a PCR amplification reaction. The data points include a cycle value associated with a PCR cycle and a product value associated with a PCR product. A baseline is identified and reaction segment associated with an exponential region of the PCR reaction is also identified. The reaction segment includes a subset of the plurality of line segments having a slope not less than a steepness criterion. A reaction line is fit to the data points associated with the reaction segment. The Cq is identified by calculating the cycle value for a data point defining the intersection of the baseline and reaction line.

主权项

1. A method of identifying a quantitation cycle (Cq) for a Polymerase Chain Reaction (PCR) amplification reaction characterized by a plurality of data points each including a cycle value associated with a PCR cycle and a product value associated with a PCR product, the method comprising:
a) fitting a plurality of line segments to the data points associated with the PCR amplification reaction; b) identifying, using a processor, a baseline of the PCR amplification reaction; c) identifying a reaction segment associated with an exponential region of the PCR amplification reaction as a first subset of the plurality of line segments having a slope not less than a steepness criteria; d) fitting a reaction line to the data points associated with the reaction segment; and e) extrapolating the reaction line to intersect the baseline at the Cq.