Evaluation of reference gene suitability for quantitative expression analysis by quantitative polymerase chain reaction in the mandibular condyle of sheep

被引：13

作者：

Jiang, Xin ^{[1
,2
]}

Xue, Yang ^{[3
]}

Zhou, Hongzhi ^{[1
]}

Li, Shouhong ^{[2
]}

Zhang, Zongmin ^{[1
]}

Hou, Rui ^{[1
]}

Ding, Yuxiang ^{[1
]}

Hu, Kaijin ^{[1
]}

机构：

[1] Fourth Mil Med Univ, Sch Stomatol, Dept Oral & Maxillofacial Surg, State Key Lab Mil Stomatol, Xian 710032, Shaanxi, Peoples R China

[2] Hubei Univ Med, Dongfeng Hosp, Dept Oral & Maxillofacial Surg, Shiyan 442008, Hubei, Peoples R China

[3] Fourth Mil Med Univ, Sch Stomatol, Dept Oral Biol, State Key Lab Mil Stomatol, Xian 710032, Shaanxi, Peoples R China

来源：

MOLECULAR MEDICINE REPORTS | 2015年 / 12卷 / 04期

基金：

中国国家自然科学基金;

关键词：

reference gene; sheep; geNorm; Norm Finder; Best Keeper; comparative Delta Ct method; REAL-TIME PCR; HOUSEKEEPING GENES; RT-PCR; NORMALIZATION; SELECTION;

D O I：

10.3892/mmr.2015.4128

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

Reference genes are commonly used as a reliable approach to normalize the results of quantitative polymerase chain reaction (qPCR), and to reduce errors in the relative quantification of gene expression. Suitable reference genes belonging to numerous functional classes have been identified for various types of species and tissue. However, little is currently known regarding the most suitable reference genes for bone, specifically for the sheep mandibular condyle. Sheep are important for the study of human bone diseases, particularly for temporomandibular diseases. The present study aimed to identify a set of reference genes suitable for the normalization of qPCR data from the mandibular condyle of sheep. A total of 12 reference genes belonging to various functional classes were selected, and the expression stability of the reference genes was determined in both the normal and fractured area of the sheep mandibular condyle. Ref Finder, which integrates the following currently available computational algorithms: geNorm, Norm Finder, Best Keeper, and the comparative Delta Ct method, was used to compare and rank the candidate reference genes. The results obtained from the four methods demonstrated a similar trend: RPLI9, ACTB, and PGK1 were the most stably expressed reference genes in the sheep mandibular condyle. As determined by Ref Finder comprehensive analysis, the results of the present study suggested that RPLI9 is the most suitable reference gene for studies associated with the sheep mandibular condyle. In addition, ACTB and PGK1 may be considered suitable alternatives.

引用

页码：5633 / 5640

页数：8

共 17 条

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