Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR

TJ Kaitu'u-Lino; R Hastie; P Cannon; S Lee; O Stock; NJ Hannan; R Hiscock; S Tong

Journal article

Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR

TJ Kaitu'u-Lino, R Hastie, P Cannon, S Lee, O Stock, NJ Hannan, R Hiscock, S Tong

Placenta | Published : 2014

DOI: 10.1016/j.placenta.2014.10.003

Abstract

Measuring mRNA expression is fundamental to placental research. Ideally, mRNA transcript numbers are directly quantified. However, PCR analysis using the DDCT method relies on the stability of housekeeping genes and only reports relative expression. Digital PCR (dPCR) directly quantifies mRNA copy number and is more accurate than quantitative PCR.We quantified absolute mRNA copy number of housekeeping genes in normotensive pre-term (n = 20), severe preeclamptic (n = 11) and term (n = 12) placenta using dPCR. Whilst there was some variation, we confirm absolute mRNA copy number of GAPDH, TOP1, CYC1 and YWHAZ in placenta does not significantly alter between these cohorts, or across gestation.

University of Melbourne Researchers

Tu'uhevaha Kaitu'u-Lino Author

Roxanne Hastie Author

Ping Cannon Author

Natalie Hannan Author

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Grants

Awarded by National Health and Medical Research Council

Funding Acknowledgements

The National Health and Medical Research Council of Australia provided salary (#1062418 to T.K, #628927 to N.H, #1050765 to S.T.) and scholarship support (#1055795 to O.S.)