Comparison of methods for relative quantification of gene expression using real-time PCR
Primerjava metod za relativno kvantifikacijo genskega izražanja s PCR v realnem času

english

2012

volume 100, issue 2

molecular genetics    genes    gene expression    quantitative real-time PCR   methods 

Univerza v Ljubljani, Biotehniška fakulteta
(Obvezni izvod spletne publikacije)

Quantitative real-time PCR (qPCR) has become a widely used tool for quantifying gene expression. Several methods for relative quantification have been developed, enabling rapid and reliable detection and quantification of specific nucleic acids. These methods, based on qPCR include: the standard curve method, the efficiency calibrated method and the 2-deltadeltaCq method. Here we analyzed if these three methods generate comparable results. To evaluate their performance, we analyzed the expression of the nuclease gene MS53_0284 from Mycoplasma synoviae type strain WVU 1853 during in vitro infection of CEC-32 cells, using qPCR. As determined, all three methods generated comparable and reliable results when all necessary conditions were fulfiled. Also, the efficiency calibrated and the standard curve methods were more suitable for quantifying small differences in relative gene expression than the 2-deltadeltaCq method.

11.06.2013