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US 2003/0148332 A1Published application
Adaptive baseline algorithm for quantitative PCR
AssigneeStratagene California
InventorsRoger Taylor, Alan Ridgeway Orr
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Abstract
The invention relates to baseline subtraction algorithms developed to reduce tube-to-tube and cycle-to-cycle variabilities during real time PCR amplification. Particularly, the invention relates to algorithms for determining the threshold cycle for the first reliable detection of the amplified nucleic acid product. The invention also relates to computer programs comprising the algorithm and methods and/systems implementing the algorithm
Claims18
A method for calculating an adaptive baseline for a plurality of sample wells in a sample block cycling through cycles of a PCR reaction, said method comprising:
- (a) plotting intensity of actual optical signal observed in a well as a function of cycle number for that well to obtain a first plot;
- (b) determining a starting cycle and an ending cycle for said well, and fitting points between and including the starting and ending cycle to a line to determine the best fit between said points and said line;
- (c) using a best fit line obtained for said well as the baseline for that well;
- (d) performing steps (a)-(c) for each of said plurality of wells.
1 more element
Dependencies are inferred from the claim text.