Identification of reference genes in blood before and after entering the plateau for SYBR green RT-qPCR studies
- Published
- Accepted
- Subject Areas
- Molecular Biology, Hematology
- Keywords
- Plateau, Hypoxia, Gene expression, Expression stability, Reference gene
- Copyright
- © 2017 Xiao 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
- 2017. Identification of reference genes in blood before and after entering the plateau for SYBR green RT-qPCR studies. PeerJ Preprints 5:e2929v1 https://doi.org/10.7287/peerj.preprints.2929v1
Abstract
Background: Tibetans have lived at high altitudes for thousands of years, and they have a unique composition of physiological traits that enable them to tolerate this hypoxic environment. However, the genetic basis of these traits is still unknown. As a sensitive and highly efficient technique, RT-qPCR is widely used in gene expression analyses to provide insight into the molecular mechanisms underlying environmental changes. However, the quantitative analysis of gene expression in blood is limited by a shortage of stable reference genes for the normalization of mRNA levels. Thus, systematic approaches were used to select potential reference genes. Results: Eight candidate reference genes (GAPDH, ACTB, 18S RNA, β2-MG, PPIA, RPL13A, TBP and SDHA) from humans were selected to assess their expression levels in blood under hypoxic environments. The expression stabilities of these candidate reference genes were evaluated using BestKeeper, geNorm and NormFinder programs. Interestingly, RPL13A was selected as the ideal reference gene to normalize the target gene expression in human blood before and after moving to the plateau. Conclusion: These results indicate that different reference genes should be selected for the normalization of gene expression in blood based on the environmental setting.
Author Comment
This is a submission to PeerJ for review.