血液总DNA提取方法的优化

Optimization for Method of Total DNA Extraction of Human Blood

目的:比较不同蛋白酶K浓度对血液总DNA浓度和纯度提取的影响作用。方法:随机抽取22份新疆建设兵团第三师51团全民体检血液样本,以TIANGEN生化科技(北京)有限公司血液提取试剂盒(离心柱型)为基础,分析11种不同蛋白酶K浓度(18.0,18.4,18.6,18.8,18.9,19.0,19.1,19.2,19.4,19.6,20.0mg/m L)对250μL血液总DNA浓度和纯度提取的影响。结果:(1)在蛋白酶K浓度在18.0~20.0mg/m L范围时,随着蛋白酶K浓度的增加,所提取的DNA浓度在19.0mg/m L时达到最大,当继续增加蛋白酶K浓度时,DNA浓度呈明显下降趋势;(2)随着蛋白酶K浓度的增加,OD_(260)/OD_(280)在18.6mg/m L时达到最大,且继续增加蛋白酶K浓度,OD260/OD280呈缓慢下降趋势;(3)蛋白酶K浓度在18.8~19.0mg/m L时,所提取的DNA浓度和纯度最佳。结论:建议TIANGEN生化科技(北京)有限公司在提取250μL人血液样本时选择蛋白酶K浓度应为18.8~19.0mg/m L,既能提高血液核酸DNA的提取质量,又可降低提取人血液DNA的成本。

Objective： Effect on concentration and purity of blood total DNA with different proteinase K concentration. Methods：A total of 22 blood samples randomly choosed from the 51 st TUAN division Hospital,First Affiliated Hospital of School of Medicine, Shihezi University. All the samples were extracted with Mammal blood genome DNA extraction kit（centrifugal column type）, produced by TIANGEN biochemical technology（Beijing,China）.Eleven different coconcenrtations of proteinase K were used to analyze their effect on concentration and purity of blood total DNA,including 18.0,18.4,18.6,18.8,18.9,19.0,19.1,19.2,19.4,19.6 and 20.0mg/m L.R esults：（1）the concentration of DNA reached peak when the concentration of proteinase K was 19.0mg/m L in the range of 18.0-19.0mg/m L for proteinase K,and decreased with increasing proteinase K concentration in the range of 19.0-20.0mg/m L for proteinase K：（2）DNA purity detected with OD260/OD280 reached the maximum when the concentration of proteinase K was 18.6mg/m L,and decreased slowly with increasing the concentration of proteinase K;（3）concentration and purity of blood total DNA were optimal when the concentration of proteinase K at18.8-19.0mg/m L.Conclusion：Our findings conveyed us that concentration of proteinase K is 18.8-19.0mg/m L when extracting 250μL human blood samples based on Mammal blood genome DNA extraction kit（TIANGEN biochemical technology）, which can improve the extraction quality of blood genome DNA,but also can reduce the cost of DNA extraction.