Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2007-12

AUTHORS

Ann-Britt Nygard, Claus B Jørgensen, Susanna Cirera, Merete Fredholm

ABSTRACT

BACKGROUND: Real-time quantitative PCR (qPCR) is a method for rapid and reliable quantification of mRNA transcription. Internal standards such as reference genes are used to normalise mRNA levels between different samples for an exact comparison of mRNA transcription level. Selection of high quality reference genes is of crucial importance for the interpretation of data generated by real-time qPCR. RESULTS: In this study nine commonly used reference genes were investigated in 17 different pig tissues using real-time qPCR with SYBR green. The genes included beta-actin (ACTB), beta-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hydroxymethylbilane synthase (HMBS), hypoxanthine phosphoribosyltransferase 1 (HPRT1), ribosomal protein L4 (RPL4), succinate dehydrogenase complex subunit A (SDHA), TATA box binding protein (TPB)and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide (YWHAZ). The stability of these reference genes in different pig tissues was investigated using the geNorm application. The range of expression stability in the genes analysed was (from the most stable to the least stable): ACTB/RPL4, TBP, HPRT, HMBS, YWHAZ, SDHA, B2M and GAPDH. CONCLUSION: Expression stability varies greatly between genes. ACTB, RPL4, TPB and HPRT1 were found to have the highest stability across tissues. Based on both expression stability and expression level, our data suggest that ACTB and RPL4 are good reference genes for high abundant transcripts while TPB and HPRT1 are good reference genes for low abundant transcripts in expression studies across different pig tissues. More... »

PAGES

67

References to SciGraph publications

  • 2007-06. Porcine transcriptome analysis based on 97 non-normalized cDNA libraries and assembly of 1,021,891 expressed sequence tags in GENOME BIOLOGY
  • 2006-12. Development of a new set of reference genes for normalization of real-time RT-PCR data of porcine backfat and longissimus dorsi muscle, and evaluation with PPARGC1A in BMC BIOTECHNOLOGY
  • 2006-12. Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR in BMC MOLECULAR BIOLOGY
  • 2005-12. Gene expression studies in prostate cancer tissue: which reference gene should be selected for normalization? in JOURNAL OF MOLECULAR MEDICINE
  • 2007-12. Validation of reference genes for quantitative RT-PCR studies in porcine oocytes and preimplantation embryos in BMC DEVELOPMENTAL BIOLOGY
  • 2006-12. Selection of a set of reliable reference genes for quantitative real-time PCR in normal equine skin and in equine sarcoids in BMC BIOTECHNOLOGY
  • 2002-07. Differential expression of GAPDH and β-actin in growing collateral arteries in MOLECULAR AND CELLULAR BIOCHEMISTRY
  • 2005-12. Selection of ovine housekeeping genes for normalisation by real-time RT-PCR; analysis of PrPgene expression and genetic susceptibility to scrapie in BMC VETERINARY RESEARCH
  • 2002-06. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes in GENOME BIOLOGY
  • 2005-12. Selection of reference genes for quantitative real-time PCR in bovine preimplantation embryos in BMC DEVELOPMENTAL BIOLOGY
  • 2005-12. Selection of reference genes for gene expression studies in human neutrophils by real-time PCR in BMC MOLECULAR BIOLOGY
  • 2005-01. Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes in LABORATORY INVESTIGATION
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    http://scigraph.springernature.com/pub.10.1186/1471-2199-8-67

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    http://dx.doi.org/10.1186/1471-2199-8-67

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/17697375


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