The preparation of PSD93 TaqMan probe for real-time fluorescence quantitative PCR

Objective:To obtain the probe of postsynaptic density 93(PSD93) for real-time fluorescence quantitative PCR(FQ-PCR).Methods:The partial CDS of PSD93 was amplified by RT-PCR from 1d postnatal SD rat brain.RT-PCR product was ligated into pGEM-T vector,and the DNA sequence was detected.The standard curve was drawn with the recombinant plasmids and the PSD93 TaqMan probe.Results:The product of RT-PCR was 113 bp which matched the size of the purpose.This DNA sequence was 100% homogeneous with the PSD93 cDNA reported. The correlation coefficient of the standard curve was above 0.99,the reaction efficiency was 0.95 and all the variation coefficients were below 20%.Conclusion:The PSD93 TaqMan probe for FQ-PCR has been successfully obtained with RT-PCR and T vector techniques.