Theory in Biosciences

, Volume 131, Issue 4, pp 215–223 | Cite as

Reference genes for measuring mRNA expression

Review

Abstract

The aim of this review is to find answers to some of the questions surrounding reference genes and their reliability for quantitative experiments. Reference genes are assumed to be at a constant expression level, over a range of conditions such as temperature. These genes, such as GADPH and beta-actin, are used extensively for gene expression studies using techniques like quantitative PCR. There have been several studies carried out on identifying reference genes. However, a lot of evidence indicates issues to the general suitability of these genes. Recent studies had shown that different factors, including the environment and methods, play an important role in changing the expression levels of the reference genes. Thus, we conclude that there is no reference gene that can deemed suitable for all the experimental conditions. In addition, we believe that every experiment will require the scientific evaluation and selection of the best candidate gene for use as a reference gene to obtain reliable scientific results.

Keywords

Reliability Beta-actin Reference genes Polymerase chain reactions 

Abbreviations

GADPH

Glyceraldehyde-3-phosphate dehydrogenase

PCR

Polymerase chain reaction

MARK3

Microtubule affinity regulating kinase 3

B2M

β2 Microglobulin

CorNV

Corneal neovascularisation

ACTB

Beta-actin

TBP

TATA binding protein

qRT-PCR

Quantitative real-time polymerase chain reaction

ALG9

Asparagine-linked glycosylation 9

RPL13A

Ribosomal protein L13a

qRT-PCR

Quantitative real-time PCR

HMBS

Hydroxymethylbilane synthase

qRT – PCR

Quantitative reverse transcription-polymerase chain reaction

HKG

Housekeeping genes

HPRT1

Hypoxanthine phosphoribosyltransferase 1

RT-qPCR

Reverse transcription-quantitative polymerase chain reaction

RT-PCR

Real time polymerase chain reaction

SAGE

Sequential analysis of gene expression

SDPH

Succinate dehydrogenase

ANOVA

One way analysis of variance

RPA

Ribonuclease protection assays

Notes

Conflict of interest

There are no competing interests cited by the authors of this paper.

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Edencore TechnologiesThaneIndia
  2. 2.Department of ZoologyThe University of MelbourneMelbourneAustralia

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