Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas)

Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan
Department of Biology, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan
Graduate Institute of Marine Biology, National Dong Hwa University, Pingtung, Taiwan
DOI
10.7287/peerj.preprints.1753v1
Subject Areas
Marine Biology, Molecular Biology
Keywords
beluga, housekeeping, quantitative PCR, gene expression
Copyright
© 2016 Chen et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ PrePrints) and either DOI or URL of the article must be cited.
Cite this article
Chen I, Wang J, Chou S, Wu Y, Li T, Leu M, Chang W, Yang WC. 2016. Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas) PeerJ PrePrints 4:e1753v1

Abstract

Reverse transcription-quantitative PCR (RT-qPCR) is used for research in gene expression, and it is vital to choose appropriate housekeeping genes (HKGs) as reference genes to obtain correct result. The purpose of this study is to determine stably expressed HKGs in blood of beluga whales (Delphinapterus leucas) that can be the appropriate reference genes in relative quantification in gene expression research. Sixty blood samples were taken from 4 beluga whales. Thirteen candidate HKGs (ACTB, B2M, GAPDH, HPRT1, LDHB, PGK1, RPL4, RPL8, RPL18, RPS9, RPS18, TFRC, YWHAZ) were tested using RT-qPCR. The stability values of the HKGs were determined by four different algorithms. Comprehensive analysis of the results revealed that RPL4, PGK1 and ACTB are strongly recommended for use in future RT-qPCR studies in beluga blood samples. This research provides recommendation of reference gene selection, which may contribute to further mRNA relative quantification research in the peripheral blood leukocytes in captive cetaceans. The gene expression assessment of the immune components in blood have potential to serve as important approach to evaluating cetacean health influenced by environmental insults.

Author Comment

This is a submission to PeerJ for review.

Supplemental Information

Raw data of Ct values in qPCR, 60 samples

DOI: 10.7287/peerj.preprints.1753v1/supp-1

Raw data of Ct values in qPCR, 30 samples

DOI: 10.7287/peerj.preprints.1753v1/supp-2