Relative quantification analysis of multi-parametric pcr experiments
OwnerIngrid Bechler, Gregor Sagnerinventors
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Abstract
An analysis method for quantitative PCR experiments including multiple genes of interest, multiple samples and multiple conditions is provided. For calculating the relative expression status of multiple target genes from multiple samples under multiple different conditions, a calculation method based on up to three different parameters is performed. In addition to normalization against one or multiple reference gene(s) and normalization against a reference sample (calibrator), a third normalization step against a base value is performed in a target-, sample- and condition-specific manner.
Claims12
A method for analysis of a polymerase chain reaction (PCR) experiment having a plurality of experimental conditions and a plurality of samples, each sample comprising one or more target nucleic acids and one or more reference nucleic acids, the method comprising
- performing a plurality of PCR amplifications of the target nucleic acids and the reference nucleic acids under the plurality of experimental conditions,
- performing a first normalization step for each experimental condition by comparing each target nucleic acid amplification to the amplification of the reference nucleic acids to obtain relative target/reference ratios,
- performing a second normalization step by comparing each relative target/reference ratio calculated in the first normalization step to a relative target/reference ratio obtained at the experimental base condition to obtain scaled target/reference ratios, and
- analyzing the PCR experiment by comparing the scaled target/reference ratios calculated in the second normalization step in a target-, sample- and condition-specific manner.
Dependencies are inferred from the claim text.