摘要
[目的]对组织DNA提取方法进行改进,建立一种从绵羊凝血块中提取基因组DNA的方法。[方法]将绵羊凝血块用眼科剪剪碎,用组织DNA抽提液裂解细胞,用蛋白酶K消化后,经过酚/氯仿抽提,无水乙醇沉淀获得基因组DNA。[结果]提取的DNA浓度为(159.90±0.70)ng/μl,A260/A280比值为1.80±0.01,分子完整,结果理想。以从凝血块中提取的DNA为模板,对绵羊BM203微卫星位点进行了PCR扩增,扩增效果良好。[结论]该方法简单、实用,提取的DNA可满足后续相关研究对DNA质量的要求,值得推广借鉴。
[ Objective ] The study aimed to establish the method of extracting genomie DNA from sheep blood clot on the basis of the improvement of method for extracting genomic DNA from tissues. [ Method ] The genomic DNA with complete primary structure and high purity was obtained from the sheep blood clot after the steps of cutting the sheep blood clot with ophthalmic scissors, cell lysis with tissue DNA extracts and digested by proteinase K, extracting with phenol/chloroform and precipitating with ethanol were performed. [ Result ] The concentration of the extracted DNA was （ 159.90 ± 0.70） ng/μl and the ratio of the A260/A280 was 1.80 ± 0.01. The sheep microsatellite locus of BM203 was amplified by using the extracted DNA from the sheep blood clot as template of PCR,and the PCR result was perfect. [ Conclusion] This method is simple and feasible, the quantity and quality of the extracted DNA can satisfy the demands for the subsequent researches. It is worth to extending and using for reference.
出处
《安徽农业科学》
CAS
北大核心
2009年第34期16771-16772,共2页
Journal of Anhui Agricultural Sciences
基金
山西省自然科学基金项目(2007011081)
山西省归国留学人员项目(2007066)
农业科技成果转化资金项目(2008GB2A300032)
关键词
绵羊凝血块
酚/氯仿抽提法
DNA提取
Sheep blood clot
Phenol/chloroform extracting method
DNA extraction
