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


CONCLUSIONS
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.
Zhang
Ning-Yuan
NY
Reproductive Medicine Center, Drum Tower Hospital Affiliated to Nanjing University School of Medicine, Nanjing, Jiangsu 210008, China.
Hu
Ya-Li
YL
Sun
Hai-Xiang
HX
Xu
Zhi-Peng
ZP
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins / Q000235:genetics*
D012333:RNA, Messenger; C000624653:TSC2 protein, human; C000624654:Tsc2 protein, mouse; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins