Selection of reference genes for gene expression studies in porcine skeletal muscle using SYBR green qPCR
Selection of reference genes for gene expression studies in porcine skeletal muscle using SYBR green qPCR
复制标题
使用 SYBR green qPCR 选择猪骨骼肌基因表达研究的参考基因
DOI：
10.1016/j.jbiotec.2010.09.949
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发表时间：
2010-11-01
影响因子：
4.1
通讯作者：
Ren, Zhuqing
中科院分区：
文献类型：
--
作者：
Feng, Xiaoting;Xiong, Yuanzhu;Ren, Zhuqing
定量PCR（qPCR）是一种快速可靠的mRNA定量方法。内部对照如参考基因用于标准化不同样品之间的mRNA水平，以精确比较基因转录水平。然而，这些参考基因的表达水平可能因细胞类型、发育阶段、物种和实验条件而异，因此适当的标准化策略是可靠结论的重要前提。本研究采用SYBR绿色qPCR技术，对猪骨骼肌中10个常用的参考基因进行了检测。我们使用geNorm和Normalt分析表达稳定性，发现PPIA、HPRT和eEF-1 γ是猪骨骼肌的合适内部对照。PPIA、HPRT和SDHA适用于西方猪的骨骼肌，PPIA、eEF-1 γ和HPRT适用于中国猪的骨骼肌。将ROCK 2的标准化qPCR数据与微阵列数据进行比较，以评估我们开发的参考基因组。(C)2010 Elsevier B. V.保留所有权利。
Quantitative PCR (qPCR) is a method for rapid and reliable quantification of mRNA. Internal controls such as reference genes are used to normalize mRNA levels between different samples for an exact comparison of gene transcription level. However, the expression levels of these reference genes may vary between cell types, developmental stages, species and experimental conditions, thus proper normalization strategy is an important precondition for reliable conclusions. In this study, we explored 10 commonly used reference genes in porcine skeletal muscle using SYBR green qPCR. We used both geNorm and NormFinder to analyze the expression stability and found that PPIA, HPRT and eEF-1 gamma were suitable internal controls for porcine skeletal muscle. However, PPIA, HPRT and SDHA were suitable for skeletal muscle of western pigs while PPIA, eEF-1 gamma and HPRT for indigenous Chinese pigs. Normalized qPCR data of ROCK2 were compared with microarray data to evaluate our developed set of reference genes. (C) 2010 Elsevier B.V. All rights reserved.