تجزیه وتحلیل داده های Quantitative Real-time Polymerase Chain Reaction در بیان ژن

Real-time PCR is broadly used as a gold standard method with high efficiency to quantify gene expression (mRNA), however, it is crucial for researchers to understand the analysis procedure in their framework in order to be able to generate reliable data. In general, there are two different methods for data analysis of RT-qPCR, including absolute quantification (AQ) and relative quantification (RQ) which will be reviewed in this article. The absolute quantification determines the input copy numbers of transcript of interest relating the PCR signals to a standard curve .The relative method, on the other hand describes the changes in gene expression of target gene relative to the control groups. The 2_ΔΔCT method is a convenient way to analyze the relative changes in gene expression from RT-qPCR experiments. The purpose of this article is to review the most common approaches of RT-qPCR. Furthermore, we will discuss the derivation and application of the 2_ΔΔCT method.

کلیدواژه ها:

Absolute Quantification ، Relative Quantification ، The 2_ΔΔCT methods ، Crossing Threshold ، Melting Curve