摘要
目的寻找最佳的从血液尤其是凝血中提取DNA的方法,减少实验和临床检测血液的用量。方法静脉抽取30份健康体检者的血样,分别抗凝、不抗凝处理后,用经优化的不需要酶和有机溶剂的抽提程序提取DNA,通过电泳和PCR进行检测。结果凝血和抗凝血的DNA产量分别是(40·2±8·86)mgDNA/L和(39·1±10·2)mgDNA/L;纯度分别为1·87±0·11和1·92±0·12。所有样本的DNA分子质量都很高,从两者的DNA样本中能很容易地扩增出t-PA基因的第8内含子区ALU等位基因二态性,因此所提取的DNA是完整可靠的。结论该方法能快速、简单、有效、无毒地从新鲜血液和凝血中提取DNA,适合于临床检测和分子生物学研究。
Objective To find an ideal method of DNA isolation from blood and especially from clotted blood and to minimize the volume of blood collected for laboratory and clinical tests. Methods DNAs were isolated from antiagglutinated and agglutinated blood samples from auricular veins of 30 healthy subjects. The DNAs of these samples were obtained by a nonenzymatic, nontoxic procedure optimized by us and determinated by agarose gel electrophoesis and PCR. Results The yields of DNA isolated from clotted blood and antiagglutinated blood were （40. 2 ± 8.86）mg DNA/L and （39. 1±10. 2）mg DNA/L, and purities were 1.87 ±0. 11 and 1.92 ± 0. 12. The DNAs that we isolated from all samples had high molecular weight and by PCR the dimorphism of ALU alleles of the 8th intron of t-PA was easy to be obtained, so they were complete and reliable. Conclusion This method is rapid, easy, efficient and nontoxic for isolation of DNA from clotted and fresh blood and meets requirements for clinical testing and molecular biology study.
出处
《基础医学与临床》
CSCD
北大核心
2007年第8期918-920,共3页
Basic and Clinical Medicine
