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Meiotic Spindle Assessment in Mouse Oocytes by siRNA-mediated Silencing
Published on: October 11, 2015
[Establishment of a method for quantifying tuberous sclerosis complex 2 mRNA in mouse single oocyte and
Ning-Yuan Zhang1, Ya-Li Hu, Hai-Xiang Sun
1Reproductive Medicine Center, Drum Tower Hospital Affiliated to Nanjing University School of Medicine, Nanjing, Jiangsu 210008, China.
Objective:
To develop a method for quantifying gene expressions in the mouse single oocyte and preimplantation embryo.
Methods:
We 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).
Results:
We 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).
Conclusion:
The 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.
