摘要
目的通过与酚/氯仿法比较,建立一种快速、经济、高效从人血凝块提取基因组DNA的简易方法。方法采用双蒸水低渗破碎红细胞,碘化钾直接、快速裂解白细胞及其核膜,氯仿/异戊醇沉淀蛋白质及残存细胞碎片,最后经异丙醇和乙醇沉淀基因组DNA。结果采用该法提取的基因组DNA浓度为(46.4±8.8)mg/L,吸光度值A260/A280为(1.79±0.23),与酚/氯仿法(46.1±8.3,1.78±0.22)比较差异无统计学意义(P>0.05);2种方法提取基因组DNA凝胶电泳条带完整,PCR扩增目的条带完整,能够满足分子生物学实验要求。结论碘化钾法是一种快速、简便、经济、高效提取人血凝块基因组DNA的方法,可以广泛运用于大规模人群基因组学研究。
Objective To establish a simple,quick and economical method for genomic DNA extraction from human blood clotting compared with traditional phenol-chloroform method. Methods Double distilled water was used to lyse red blood cells（RBC） and saturated potassium iodide to lyse white blood cells（WBC） and its nuclear membrane.Chloroform and isoamyl alcohol was used to precipitate proteins and the residues of cells.The the genomic DNA was precipitated by isopropanol and ethanol. Results The average quantity of genomic DNA extracted was 46.4±8.8mg/L and the ratio of A260/A280 was 1.79±0.23.Furthermore,no statistical signifcance was observed when compared with phenol-chloroform method.The electrophoretic bands of genomic DNA and PCR amplification products were distinct,and the extracted genomic DNA reached the standard for molecular biological experiment. Conclusion The postassium iodide method is a simple,quick,economical and efficient way for genomic DNA extraction from human blood clotting,and could be used in large genomics study.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2011年第4期519-520,共2页
Chinese Journal of Public Health
基金
广东省科技计划项目(2008B030301100)
中国营养学会营养科学基金(2007)
关键词
人血凝块
DNA提取
碘化钾
human blood clotting
DNA extraction
postassium iodide
