This paper presents a comprehensive algorithm for quantifying real-time polymerase chain reaction (qRT-PCR) data without the need for a standard curve or subjective judgments. The algorithm, called Real-time PCR Miner, uses a four-parameter logistic model to fit raw fluorescence data and identify the exponential phase of the reaction. It then applies a three-parameter simple exponent model to estimate amplification efficiency and the cycle threshold (CT) using iterative nonlinear regression and weighted averages. The CT is determined by the first positive second derivative maximum of the logistic model. The algorithm is noise-resistant and independent of the PCR platform used. It provides objective quantification of qRT-PCR results by directly calculating efficiency and CT from individual PCR reactions. The method was validated using data from multiple real-time PCR platforms and showed high accuracy and consistency compared to standard curve methods. The algorithm also accounts for noise in the fluorescent signal during exponential amplification and provides more reliable efficiency estimates. The results demonstrate that the algorithm produces accurate and consistent quantification of qRT-PCR data, making it a valuable tool for researchers in biomedical and molecular biology fields.This paper presents a comprehensive algorithm for quantifying real-time polymerase chain reaction (qRT-PCR) data without the need for a standard curve or subjective judgments. The algorithm, called Real-time PCR Miner, uses a four-parameter logistic model to fit raw fluorescence data and identify the exponential phase of the reaction. It then applies a three-parameter simple exponent model to estimate amplification efficiency and the cycle threshold (CT) using iterative nonlinear regression and weighted averages. The CT is determined by the first positive second derivative maximum of the logistic model. The algorithm is noise-resistant and independent of the PCR platform used. It provides objective quantification of qRT-PCR results by directly calculating efficiency and CT from individual PCR reactions. The method was validated using data from multiple real-time PCR platforms and showed high accuracy and consistency compared to standard curve methods. The algorithm also accounts for noise in the fluorescent signal during exponential amplification and provides more reliable efficiency estimates. The results demonstrate that the algorithm produces accurate and consistent quantification of qRT-PCR data, making it a valuable tool for researchers in biomedical and molecular biology fields.