The Preparation Of Satb2 Gene Standard Plasmids For Real-time Quantitative PCR And Its Application

[objective] To prepare the Satb2 gene standard plasmids for real-time quantitative PCR.[Methods] The total RNA was isolated from the maxillary process of the mouse embryo,and was used to synthesize the first-strand cDNA via reverse transcription. With Satb2 gene specific PCR and gel purification. The Satb2 gene fragments were obtained. The fragment was then inserted into the pGMT plasmid vector. After massive extraction and identification ,the plasmid were quantified and the standard curve was established. At last,the standard curve was applied in the real-time quantitative PCR.[Results] The amplified products of the recombinant plasmid by PCR and gene sequencing confirmed that the pGMT-Satb2 gene was successfully cloned.[Conclusions] The recombinant plasmid and standard curve to detect the Satb2 gene by the"Taqman"approach was good at sensitivity, specificity, quantification and linear function. It can be a standard method of real-time qPCR for detection of Satb2 gene.