天冬药材基因组DNA提取方法研究
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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条带。[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。
[Objective]The research aimed to extract high-quality 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.
[Objective]The research aimed to extract high-quality 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.
引文
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[6] 罗焜,马培,姚辉,等.中药DNA条形码鉴定中的DNA提取方法研究[J].世界科学技术(中医药现代化),2012,14(2):1433-1439.
[7] 符德欢,朱高倩,郭佳玉,等.改良CTAB法提取重楼属3种药用植物干燥根茎DNA[J].中药材,2017,40(6):1295-1299.
[8] 苟惠,王译伟,郑茜,等.含当归中成药的DNA提取及其分子鉴定[J].中国实验方剂学杂志,2018(1):44-50.
[9] 纪宝玉,裴莉昕,陈随清,等.山茱萸药材鲜品与干品总 DNA 提取方法比较[J].中国实验方剂学杂志,2015,21(10):70-72.
[10] JIANG J,YANG C P.Molecular biology experiment principle and technology [M].Haerbin:Northeast Forestry University Publishing House,2000.
[11] 袁天翊,阎雨,方莲花,等.利用 RAPD 法为中药材标准物质建立DNA图谱[J].中国新药杂志,2013,22(19):2250-2255.
[12] 姜颖,杨欣,于英君.异地板蓝根基因组DNA指纹图谱建立及RAPD-PCR 反应体系优化[J].中医药学报,2014,42(5):64-66.
[2] 林钰文.中药天冬研究进展[J].海峡药学,2008,20(6):90-93.
[3] 国家中医药管理局.中医病证诊断疗效标准:ZY/T 001.1～001.9—94[S].南京:南京大学出版社,1994:36-37.
[4] 国家药典委员会.中华人民共和国药典[S].北京:中国医药科技出版社,2015:55-56.
[5] 沈阳,陈海生,王琼.天冬化学成分的研究(Ⅱ)[J].第二军医大学学报,2007,28(11):1241-1244.
[6] 罗焜,马培,姚辉,等.中药DNA条形码鉴定中的DNA提取方法研究[J].世界科学技术(中医药现代化),2012,14(2):1433-1439.
[7] 符德欢,朱高倩,郭佳玉,等.改良CTAB法提取重楼属3种药用植物干燥根茎DNA[J].中药材,2017,40(6):1295-1299.
[8] 苟惠,王译伟,郑茜,等.含当归中成药的DNA提取及其分子鉴定[J].中国实验方剂学杂志,2018(1):44-50.
[9] 纪宝玉,裴莉昕,陈随清,等.山茱萸药材鲜品与干品总 DNA 提取方法比较[J].中国实验方剂学杂志,2015,21(10):70-72.
[10] JIANG J,YANG C P.Molecular biology experiment principle and technology [M].Haerbin:Northeast Forestry University Publishing House,2000.
[11] 袁天翊,阎雨,方莲花,等.利用 RAPD 法为中药材标准物质建立DNA图谱[J].中国新药杂志,2013,22(19):2250-2255.
[12] 姜颖,杨欣,于英君.异地板蓝根基因组DNA指纹图谱建立及RAPD-PCR 反应体系优化[J].中医药学报,2014,42(5):64-66.