摘要
目的从血液中分离基因组DNA是临床诊断和基因组研究的基础,血液中的众多指标检测对于很多疾病的早期诊断、预后判断、复发监测及疗效评估都有重要的临床意义。因此需要改善血液基因组DNA提取方法,提高实验效率。方法本方法以tween-20结合盐酸胍的方法释放DNA,无需蛋白酶K消化,通过自制滤膜吸附柱,可以在15 min内得到高纯度的DNA,对冻存及各种抗凝剂处理的血液样本均适用。结果 200μl的血液可以获得6μg左右的基因组DNA,A260/A280>1.8,浓度为30 ng/μl,高于一般商业化试剂盒的血液基因组DNA提取浓度(10 ng/μl)。PCR结果表明产物无抑制剂残留。结论该方法不仅能大大缩短DNA提取操作时间,适用于自动化提取操作,还能节约费用,可广泛使用与临床分子诊断和科研。
Objective Blood genomic DNA is critical for early diagnosis,prognosis and curative effect evaluation of many clinical diseases. So a fast and novel method for extraction of genomic DNA from blood is developed to improve the experiment efficiency. Methods Guanidine hydrochloride containing 10% tween-20 was used to extract DNA without proteinase K digestion. Then the pure DNA can be obtained with self-made filter membrane adsorption column only in 15 min,and it was suitable for both fresh and frozen human blood. Results About 6 μg of genomic DNA were obtained from 200 μl blood sample with an A260 /280ratio above 1. 8 and a concentration of 30 ng / μl,higher than the extraction concentration of general commercial kit（ 10 ng/μl）. PCR experiments indicated that the final DNA extraction contained no inhibitory substances. Conclusion This novel method with low cost and timesaving makes it popular for clinical molecular diagnosis and research.
出处
《中国卫生检验杂志》
北大核心
2014年第7期982-984,共3页
Chinese Journal of Health Laboratory Technology
基金
宁波市科技创新团队项目(2011B82016)
