Optimization of β-actin gene as a housekeeping gene for quantification of mRNA levels of target genes in buffalo endometrium

Abstract

In order to improve (re)production, the tissue specific expression profile of gene(s) involved in the various stages of reproduction should be studied thoroughly. For the same, Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis of target genes in various biological samples could fulfil the aim. Various published literatures are evidence that the qRT-PCR technique is one of the most widely used and practical methods for detecting gene expression levels. In this technique, the use of housekeeping or reference gene acts as a calibrator, plays a vital role for quantification of mRNA expression levels of target gene in order to achieve objective and reliable findings. Housekeeping genes are responsible for the maintenance of basic cellular activities that are required for a cell's existence, regardless of the cell's specialised role in the tissue or organism. However, designing of primers and optimization of PCR conditions for a housekeeping gene was required to study the function of various other genes in buffalo endometrium. Therefore, the present study was aimed to optimize the β-actin gene as a housekeeping gene that stably expresses in the buffalo uterine endometrium, and establish β-actin gene primers for qRT-PCR technique.