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

ISRN biotechnology Pub Date : 2013-01-28 eCollection Date: 2013-01-01 DOI:10.5402/2013/735053

Neha Kapila, Amit Kishore, Monika Sodhi, Ankita Sharma, Pawan Kumar, A K Mohanty, Tanushri Jerath, M Mukesh

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引用次数: 25

Abstract

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.

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水牛热应激乳腺上皮细胞qRT-PCR分析内参基因的鉴定

基因表达研究需要适当的归一化方法来正确评估内参基因。迄今为止，关于确定水牛中合适的内参基因的研究报告并不多。本研究旨在确定热应激水牛乳腺上皮细胞(MECs)中合适的内参基因。简单地说，将水牛乳腺MEC培养物在42°C下暴露1小时，然后在37°C下恢复不同的时间间隔(从30 m到48 h)。采用geNorm、NormFinder和BestKeeper三种不同的算法对来自不同功能类别的16个潜在内参基因进行稳定性评估。我们的数据确定RPL4、EEF1A1和RPS23基因是最合适的内参基因，可以用于热应激水牛mec qPCR数据的规范化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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ISRN biotechnology

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