| 引用本文: | 詹羽姣,盛萍,张鹏葛,安露莎,张煊.贝母属药用贝母总DNA提取方法的比较研究[J].中国现代应用药学,2015,32(8):915-919. |
| ZHAN Yujiao,SHENG Ping,ZHANG Pengge,AN Lusha,ZHANG Xuan.Comparative Research on Genomic DNA Extraction Methods from Medicinal Fritillaria L.[J].Chin J Mod Appl Pharm(中国现代应用药学),2015,32(8):915-919. |
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| 摘要: |
| 目的 比较筛选最适于贝母属药用贝母的总DNA提取方法。方法 利用试剂盒法、改良的CTAB法、改良的SDS法提取贝母属药用贝母的总DNA；通过核酸蛋白检测仪及琼脂糖凝胶电泳方法检测其浓度和纯度；进行ISSR-PCR扩增检测所得总DNA质量。结果 3种方法均可提取出产量较高的基因组DNA，但试剂盒法相较提取的总DNA纯度最高，质量最好，并且均能扩增出丰富清晰的条带，稳定性高，操作简单耗时短。改良的CTAB法、SDS法提取的总DNA纯度、质量均次于试剂盒法，操作复杂耗时长，不适用于下游的分子生物学实验。结论 试剂盒法为贝母属药用贝母总DNA提取的最佳方法，其所得样品适用于PCR及其他分子生物学研究。 |
| 关键词: 贝母属 总DNA提取 ISSR |
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| 基金项目:国家自然科学基金(81160544) |
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| Comparative Research on Genomic DNA Extraction Methods from Medicinal Fritillaria L. |
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ZHAN Yujiao1, SHENG Ping1,2, ZHANG Pengge1, AN Lusha1, ZHANG Xuan1
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1.School of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi 830011, China;2.Xinjiang Key Laboratory of Famous Prescription and Science of Formulas, Urumqi 830011, China
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| Abstract: |
| OBJECTIVE To choose the most suitable extraction method of total DNA in medicinal Fritillaria L.. METHODS Using the kit method, modified CTAB method, modified SDS method total DNA of Fritillaria L. were extracted; the potency and purity of the total DNA were detected by nucleic acid protein detector and agarose gel electrophoresis method; ISSR-PCR amplification detection obtained DNA quality. RESULTS higher yield of genomic DNA can be extracted from the plant Fritillaria L. by 3 kinds of methods. But compared with the total DNA, kit method was the highest, best quality of the extracted, it could amplify the bands rich and clear, and high stability, simple operation and short time. The purity and quality of total DNA extracted by modified CTAB method or SDS method were inferior those by kit method. Moreover, CTAB method or SDS method was complex and time-consuming operation, not for downstream molecular biology experiments. CONCLUSION The kit method is the best way to extract total DNA from medicinal plants of Fritillaria L., and can be used directly in PCR and other biological research. |
| Key words: Fritillaria L. total DNA extraction ISSR |
