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- Title
Selection and evaluation of new reference genes for RT-qPCR analysis in Epinephelus akaara based on transcriptome data.
- Authors
Wang, Huan; Zhang, Xiang; Liu, Qiaohong; Liu, Xiaochun; Ding, Shaoxiong
- Abstract
Groupers are an economically important fish species in world fishery markets. Because many studies using RT-qPCR have addressed gene expression in groupers, appropriate reference genes are required to obtain reliable and accurate results. In this study, the most suitable reference genes were identified from eleven candidate genes of one of the most valuable species, Epinephelus akaara, in a range of different experimental conditions. Using the software packages geNorm, NormFinder, BestKeeper and refFinder, three traditionally used reference genes, β-actin (β-ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-2-microglobulin (B2M), were identified as not suitable for E. akaara gene expression studies, whereas two newly identified reference genes, conserved oligomeric Golgi complex subunit 5 (Cog5) and brefeldin a-inhibited guanine nucleotide-exchange protein 1 (ARFGEF1), could be universally applied under all the tested conditions. These data provide the foundation for more precise results in RT-qPCR studies of gene expression in E. akaara and other Epinephelus species.
- Subjects
POLYMERASE chain reaction; EPINEPHELUS akaara; GENETIC transcription; GLYCERALDEHYDEPHOSPHATE dehydrogenase; BETA 2-microglobulin
- Publication
PLoS ONE, 2017, Vol 12, Issue 2, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0171646