National Journal of Andrology 2009;15(5):417-421

Establishment of a method for quantifying tuberous sclerosis complex 2 mRNA in mouse single oocyte and preimplantation embryo.

Ning-Yuan ZHANG 1 ; Ya-Li HU ; Hai-Xiang SUN ; Zhi-Peng XU

Affiliations

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Country

China

Language

Chinese

Abstract

OBJECTIVETo develop a method for quantifying gene expressions in the mouse single oocyte and preimplantation embryo.

METHODSWe quantified the message RNA (mRNA) expression of the TSC2 gene in the single oocyte and preimplantation embryo by capillary electrophoresis using the exogenic mutation TSC2 gene as the reference and amplification by competition polymerase chain reaction (PCR).

RESULTSWe successfully established the method for quantifying the mRNA expression of the TSC2 gene, with good linear relations between the mRNA level of the TSC2 gene and the dilution degree of the reference gene (r = -0.987).

CONCLUSIONThe level of the mouse TSC2 gene expression can be effectively quantified by competition PCR and capillary electrophoresis, which has provided a molecular base for evaluating the quality of human oocytes and preimplantation embryos.