| 张国林,魏星,邢以文,等.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[J].安徽医药,2021,25(10):1946-1949. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2021.10.009 |
| 中文关键词: 贝母属 主成分分析 聚合酶链式反应 限制性片段长度多态性 川贝母 平贝母 复方川贝散 |
| 英文关键词: Fritillaria Principal component analysis Polymerase chain reaction Restriction fragment length polymorphism Fritillariae cirrhosae bulbus Fritillarae ussuriensis bulbus Fufang chuanbei san |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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 SmaⅠ restriction 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 SmaⅠ 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. |
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