天冬药材基因组DNA提取方法研究
摘要 [目的]从天冬药材中提取可用于RAPD扩增的高质量基因组DNA。[方法]分别采用改良CTAB法、SDS法以及植物基因组DNA提取试剂盒法提取天冬药材基因组DNA;通过电泳、DNA浓度与纯度检测以及RAPD扩增效果确定天冬药材基因组DNA的最佳提取方法。[结果]CTAB法提取的基因组DNA较完整,DNA浓度与纯度均较高,可获得丰富的、清晰的RAPD條带。[结论]CTAB法为天冬药材基因组DNA提取的最佳方法,提取的DNA可用于天冬RAPD分析,该方法可为其他药材基因组DNA提取提供参考。
关键词 天冬;药材;基因组DNA提取;RAPD
中图分类号 S567.23 文献标识码 A
文章编号 0517-6611(2019)08-0171-03
doi:10.3969/j.issn.0517-6611.2019.08.045
Abstract [Objective]The research aimed to extract highquality genomic DNA which was used for RAPD amplification from herbal pieces such as ASPARAGI RADIX. [Method]Different methods, CTAB method, SDS method and DNA secure Plant Kit method were used to genomic DNA extract from ASPARAGI RADIX.The DNA samples obtained by the above methods were tested by agarose gel electrophoresis, concentration and purity of DNA and RAPD amplification. [Result]The more profuse and clear RAPD bands were obtained with random primers S7 by using improved CTAB. [Conclusion]CTAB method and RAPD reaction system can be used to RAPD analysis in ASPARAGI RADIX,which can provide certain reference for the extraction of genomic DNA of Chinese traditional medicine.
Key words ASPARAGI RADIX;Medicinal materials;Extraction of genomic DNA;RAPD
天冬(ASPARAGI RADIX)为百合科植物天冬Asparagus cochinchinensis (Lour.) Merr.的干燥块根,主要含有多糖、皂苷等成分[1-3],具有养阴润燥、清肺生津等功效,用于肺燥干咳、内热消渴、肠燥便秘、热病津伤等[4],为乌鸡白凤丸、二冬膏、党参固本丸、冬白梅片、咽炎片等著名制剂的主要原料药材。天冬在我国分布较广泛,其主产于广西、贵州、云南等地,并以贵州产的天冬质量最好[5]。
近年来,DNA分子标记鉴定技术相对于传统经验鉴别...
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