Transformation of real-time PCR fluorescence data to target gene quantity.

Gene quantification has been significantly simplified by the development of the real-time polymerase chain reaction (PCR) technique, which has become the most sensitive method for detection and quantification of specific nucleic acid sequences. Quantitative real-time PCR is widely used in basic biological and medical research as well as for diagnostics and monitoring of the disease process [for reviews see 1–6]. Fluorescence data obtained from real-time PCR must be transformed by some method of real-time PCR data analysis to obtain the target DNA or RNA quantity. Two methods are widely used for the quantification—the standard curve method and the ‘‘2 ” method [7,8]. Recently, other techniques for real-time PCR data analysis have been developed and have become alternatives to the two classical methods. This review describes and compares the traditional and the newly developed techniques used for quantification of specific DNA and RNA sequences.

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