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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 rapid, economical and efficient method for extracting genomic DNA from human blood clots by comparing with phenol / chloroform method. Methods Red blood cells were infiltrated by double distilled water, potassium iodide was used to directly and rapidly lyse leucocytes and their nuclear membranes, and protein was precipitated by chloroform / isoamyl alcohol and residual cell debris. Genomic DNA was precipitated by isopropanol and ethanol. Results The concentration of genomic DNA extracted by this method was (46.4 ± 8.8) mg / L and the absorbance A260 / A280 was (1.79 ± 0.23). There was no significant difference with the phenol / chloroform method (46.1 ± 8.3, 1.78 ± 0.22) Significance (P> 0.05). The two methods extracted genomic DNA gel electrophoresis bands intact, PCR amplification of the purpose of band integrity, to meet the requirements of molecular biology experiments. Conclusion Potassium iodide is a rapid, simple, economical and efficient method for extracting genomic DNA from human blood clots. It can be widely used in large-scale population genomics research.