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

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Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. / Hillig, Ann-Britt Nygaard; Jørgensen, Claus Bøttcher; Cirera, Susanna; Fredholm, Merete.

In: BMC Molecular Biology, Vol. 8, No. 67, 2007.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Hillig, A-BN, Jørgensen, CB, Cirera, S & Fredholm, M 2007, 'Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR', BMC Molecular Biology, vol. 8, no. 67. https://doi.org/10.1186/1471-2199-8-67

APA

Hillig, A-B. N., Jørgensen, C. B., Cirera, S., & Fredholm, M. (2007). Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Molecular Biology, 8(67). https://doi.org/10.1186/1471-2199-8-67

Vancouver

Hillig A-BN, Jørgensen CB, Cirera S, Fredholm M. Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Molecular Biology. 2007;8(67). https://doi.org/10.1186/1471-2199-8-67

Author

Hillig, Ann-Britt Nygaard ; Jørgensen, Claus Bøttcher ; Cirera, Susanna ; Fredholm, Merete. / Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. In: BMC Molecular Biology. 2007 ; Vol. 8, No. 67.

Bibtex

@article{1b2b9a80a1c311ddb6ae000ea68e967b,
title = "Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR",
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 I (HPRT I), 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 HPRT I 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 HPRT I are good reference genes for low abundant transcripts in expression studies across different pig tissues.",
author = "Hillig, {Ann-Britt Nygaard} and J{\o}rgensen, {Claus B{\o}ttcher} and Susanna Cirera and Merete Fredholm",
year = "2007",
doi = "10.1186/1471-2199-8-67",
language = "English",
volume = "8",
journal = "B M C Molecular Biology",
issn = "1471-2199",
publisher = "BioMed Central Ltd.",
number = "67",

}

RIS

TY - JOUR

T1 - Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR

AU - Hillig, Ann-Britt Nygaard

AU - Jørgensen, Claus Bøttcher

AU - Cirera, Susanna

AU - Fredholm, Merete

PY - 2007

Y1 - 2007

N2 - 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 I (HPRT I), 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 HPRT I 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 HPRT I are good reference genes for low abundant transcripts in expression studies across different pig tissues.

AB - 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 I (HPRT I), 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 HPRT I 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 HPRT I are good reference genes for low abundant transcripts in expression studies across different pig tissues.

U2 - 10.1186/1471-2199-8-67

DO - 10.1186/1471-2199-8-67

M3 - Journal article

C2 - 17697375

VL - 8

JO - B M C Molecular Biology

JF - B M C Molecular Biology

SN - 1471-2199

IS - 67

ER -

ID: 8086725