聚合酶链式反应-限制性片段长度多态性分析复方川贝散中川贝母成分

Analysis of fritillariae cirrhosae bulbus in Fufang Chuanbei San by PCR-restriction fragment length polymorphism

目的采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)分析复方川贝散中川贝母成分。方法研究起止时间为2019年2月至2019年5月。新型快速植物基因组DNA提取试剂盒分别提取复方川贝母和蛇胆川贝液中DNA,PCR扩增后采用SmaⅠ限制性内切酶酶切,并进行琼脂糖凝胶电泳分析。同时,对川贝母含量为0%-40%的样本进行PCR-RFLP检测。结果复方川贝散和蛇胆川贝液PCR扩增后均出现特异性条带,但经SmaⅠ酶切后复方川贝散在100-250 bp出现两条特异性酶切条带,而蛇胆川贝液无特异性条带。川贝母含量为4%时,经PCR-RFLP分析在100-250 bp得到两条特异性DNA条带。结论PCR-RFLP可有效分析复方川贝散中川贝母成分,且对含量超过4%的样本可有效检出。

Objective To analyze fritillariae cirrhosae bulbus in Fufang Chuanbei San by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Methods The present research was conducted from February 2019 to May 2019.A new rapid plant genomic DNA extraction kit was developed to extract DNA from Fufang Chuanbei San and Shedan Chuanbei Ye.PCR amplified products were analyzed by SmaⅠrestriction enzyme digestion and agarose gel electrophoresis,and PCR-RFLP was used to detect the samples with contents of fritillariae cirrhosae bulbus from 0%to 40%.Results Specific bands were found after PCR amplification of compound Fufang Chuanbei San and Shedan Chuanbei Ye.After Sma I digestion,there were two specific bands between 100 bp and 250 bp in compound Fufang Chuanbei San,but there was no specific band in Shedan Chuanbei Ye.When the content of fritillariae cirrhosae bulbus was 4%,two specific DNA bands were obtained between 100 bp and 250 bp by PCR-RFLP analysis.Conclusion PCRRFLP can effectively analyze the components of fritillariae cirrhosae bulbus in Fufang Chuanbei San and can effectively detect samples with more than 4%of fritillariae cirrhosae bulbus.