摘要
目的:利用不同方法对白及基因组DNA进行提取,比较提取质量的差异,找到一种最适于白及基因组的DNA提取方法,以满足白及DNA条形码研究的需要。方法:采用十六烷基三甲基溴化铵(CTAB)法、十二烷基硫酸钠(SDS)法、碱裂解法和经本文改良的CTAB法提取白及叶片基因组DNA,利用琼脂糖凝胶电泳、紫外分光光度法及PCR扩增效果检测所提DNA的质量。结果:CTAB法、SDS法和改良CTAB法均能从白及中提取到基因组DNA,而碱裂解法几乎提取不到。其中改良CTAB法提取的DNA纯度和完整度均高于其他方法,PCR扩增效率达到100%,且扩增条带明亮清晰。结论:经本文改良的CTAB法是提取白及基因组DNA的最佳方法,该方法提取的基因组DNA适用于DNA条形码研究。
Objective： By comparing different methods for genomic DNA extraction, the best genomic DNA extraction method of Bletilla striata, suitable for DNA barcoding, was selected. Method： CTAB method, SDS method, alkaline lysis method and improved CTAB method were used to extract genomic DNA from B. striata. The yield and quality of genomic DNA were evaluated by Agarose gel electrophoresis, UV-spectrophotometry and PCR. Result： CTAB method, SDS method and improved CTAB method could extract genomic DNA from B. striata, while alkaline lysis method yields little. The quality of genomic DNA yielded by improved CTAB method were higher than those yielded by other three methods, and its amplification success rate was 100 % with clear electrophoretic bands of amplified products. Conclusion： The improved CTAB method was the best method for genomic DNA extraction from B. striata and could satisfy the need for DNA barcoding study.
出处
《中国实验方剂学杂志》
CAS
北大核心
2014年第12期119-123,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金(81360636)
贵州省中药现代化科技产业研究开发专项项目(黔科合中药字[2010]5027号
黔科合重G字[2013]4001)
贵阳市科学技术计划项目(筑科合同[2012204]20号)
