Quantitative Evaluation and Selection of Reference Genes for Quantitative RT-PCR in Mouse Acute Pancreatitis
Joint Authors
Tang, Chengwei
Yan, Zhaoping
Gao, Jinhang
Lv, Xiuhe
Yang, Wenjuan
Wen, Shilei
Tong, Huan
Source
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-03-16
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
The analysis of differences in gene expression is dependent on normalization using reference genes.
However, the expression of many of these reference genes, as evaluated by quantitative RT-PCR, is upregulated in acute pancreatitis, so they cannot be used as the standard for gene expression in this condition.
For this reason, we sought to identify a stable reference gene, or a suitable combination, for expression analysis in acute pancreatitis.
The expression stability of 10 reference genes (ACTB, GAPDH, 18sRNA, TUBB, B2M, HPRT1, UBC, YWHAZ, EF-1α, and RPL-13A) was analyzed using geNorm, NormFinder, and BestKeeper software and evaluated according to variations in the raw Ct values.
These reference genes were evaluated using a comprehensive method, which ranked the expression stability of these genes as follows (from most stable to least stable): RPL-13A, YWHAZ > HPRT1 > GAPDH > UBC > EF-1α > 18sRNA > B2M > TUBB > ACTB.
RPL-13A was the most suitable reference gene, and the combination of RPL-13A and YWHAZ was the most stable group of reference genes in our experiments.
The expression levels of ACTB, TUBB, and B2M were found to be significantly upregulated during acute pancreatitis, whereas the expression level of 18sRNA was downregulated.
Thus, we recommend the use of RPL-13A or a combination of RPL-13A and YWHAZ for normalization in qRT-PCR analyses of gene expression in mouse models of acute pancreatitis.
American Psychological Association (APA)
Yan, Zhaoping& Gao, Jinhang& Lv, Xiuhe& Yang, Wenjuan& Wen, Shilei& Tong, Huan…[et al.]. 2016. Quantitative Evaluation and Selection of Reference Genes for Quantitative RT-PCR in Mouse Acute Pancreatitis. BioMed Research International،Vol. 2016, no. 2016, pp.1-11.
https://search.emarefa.net/detail/BIM-1099037
Modern Language Association (MLA)
Yan, Zhaoping…[et al.]. Quantitative Evaluation and Selection of Reference Genes for Quantitative RT-PCR in Mouse Acute Pancreatitis. BioMed Research International No. 2016 (2016), pp.1-11.
https://search.emarefa.net/detail/BIM-1099037
American Medical Association (AMA)
Yan, Zhaoping& Gao, Jinhang& Lv, Xiuhe& Yang, Wenjuan& Wen, Shilei& Tong, Huan…[et al.]. Quantitative Evaluation and Selection of Reference Genes for Quantitative RT-PCR in Mouse Acute Pancreatitis. BioMed Research International. 2016. Vol. 2016, no. 2016, pp.1-11.
https://search.emarefa.net/detail/BIM-1099037
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1099037