赵昕, 张继伟, 陈国雄, 李玉霖. 荒漠植物中总脱氧核糖核酸分子的提取方法[J]. 分析测试技术与仪器, 2020, 26(1): 37-41. DOI: 10.16495/j.1006-3757.2020.01.007
引用本文: 赵昕, 张继伟, 陈国雄, 李玉霖. 荒漠植物中总脱氧核糖核酸分子的提取方法[J]. 分析测试技术与仪器, 2020, 26(1): 37-41. DOI: 10.16495/j.1006-3757.2020.01.007
ZHAO Xin, ZHANG Ji-wei, CHEN Guo-xun, LI Yu-lin. Extraction Method of Total Deoxyribo Nucleic Acid from Desert Plants[J]. Analysis and Testing Technology and Instruments, 2020, 26(1): 37-41. DOI: 10.16495/j.1006-3757.2020.01.007
Citation: ZHAO Xin, ZHANG Ji-wei, CHEN Guo-xun, LI Yu-lin. Extraction Method of Total Deoxyribo Nucleic Acid from Desert Plants[J]. Analysis and Testing Technology and Instruments, 2020, 26(1): 37-41. DOI: 10.16495/j.1006-3757.2020.01.007

荒漠植物中总脱氧核糖核酸分子的提取方法

Extraction Method of Total Deoxyribo Nucleic Acid from Desert Plants

  • 摘要: 建立了荒漠植物总脱氧核糖核酸分子(DNA)的提取方法.荒漠植物叶片加少量交联聚乙烯吡咯烷酮(PVPP粉末)研磨三次以上,得到样品超细粉末.样品粉末迅速加入前处理缓冲液,混匀后低速离心,弃上清留下沉淀物,在沉淀物中加入等体积预热的提取裂解液,混匀后60~70℃温浴1 h.高速离心提取上清液加入纯化液混匀、抽提、离心.再次提取混合液上清,加入预冷的异丙醇,-18℃沉淀DNA 0.5 h以上,异丙醇溶液高速离心,取沉淀用75%乙醇清洗两遍,晾干,加入超纯水溶解.方法具有操作简单和耗时少等优点,操作过程大约2~3 h.DNA损失少,产量高于500 ng/μL.方法提取主要荒漠植物叶片DNA纯度高、完整性好,具有广泛适用性.

     

    Abstract: An extraction method of total deoxyribo nucleic acid(DNA) from desert plants has been established. The steps of the method are as follows:add a small amount of PVPP powder to the leaves of desert plants and grind them for more than three times to get the ultra-fine powder of the sample, immediately add the pre-treatment buffer solution to the sample powder, mix them evenly and centrifuge them at low speed, then discard the supernatant and leave the precipitate, add an equal volume of preheated extraction cracking solution to the precipitate, mix them evenly on a warm bath at 60~70℃ for one hour, extract the supernatant using high-speed centrifugation. And then add the purified mixture to the solution and mix, extract and centrifuge, extract the supernatant of the mixture again, add the precooled isopropanol and precipitate DNA at -18℃ for more than half an hour, after that, centrifuge the isopropanol solution in high-speed, wash the precipitate twice with 70% ethanol, air dry and to dissolve the precipitate with ultra-pure water. The advantages of the technology are simple operation and less time-consuming, and the operation process is about 2~3 h. The yield of DNA was higher than 500 ng/μL. This method has the advantages of high purity, good integrity and wide applicability.

     

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