Table of Contents
ISRN Biotechnology
Volume 2013, Article ID 735053, 9 pages
Research Article

Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis)

1DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India
2Biotechnology Division, Singhania University, Jhunjhnu, Rajasthan 333515, India
3Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India

Received 3 November 2012; Accepted 23 November 2012

Academic Editors: R. Greiner and J. Jia

Copyright © 2013 Neha Kapila et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified RPL4, EEF1A1, and RPS23 genes to be the most appropriate reference genes that could be utilized for normalization of qPCR data in heat-stressed buffalo MECs.