人非抗凝血块中基因组DNA提取不同方法的比较

摘要

Objective:To compare three different methods in extracting genomic DNA from human non-anticoagulant blood clot. Methods:The genomic DNA was isolated from long-time cryopreserved human non-anticoagulant blood clots by using the phenol-chloro-form methods,TIANGEN kit and Invitrigen purelinkTM kit respectively. The concentration and purity of DNA were measured by ultravio-let spectrophotometer and agarose gel electrophoresis,and PCR experiment was performed using the extracted genome DNA as a tem-plate. Results:The genomic DNA was extracted successfully by three methods. The DNA concentration extracted by the three methods were 35. 56 ± 15. 27 ng/μl,45. 13 ± 16. 54 ng/μl,57. 93 ± 14. 5 ng/μl respectively and the statistical significant difference was found among the three different methods in the purity of DNA(P<0. 01). However,A260/A280 of the three groups were 1. 46 ± 0. 19,1. 49 ±0.16,1.519±0.23 and the statistical difference didn’t existed(P>0.05).The target band all appeared after PCR from genomic DNA by three extracting methods and the gray value of three group PCR products was 75. 421 ± 3. 435,149. 32 ± 6. 875 and 229. 52 ± 19. 55 and there was statistical significant difference(P<0. 01). Conclusion:The genomic DNA could be extracted from human non-anticoagulant blood clots by three different methods,however,the Invitrigen purelinkTM kit was the best.%目的：：比较三种不同方法提取人非抗凝血块基因组DNA的效果差异。方法：分别采用酚/氯仿法,天根试剂盒,Invitrigen purelinkTM试剂盒提取冻存时间较长的非抗凝血块基因组DNA,测定浓度、OD260/280值,琼脂糖凝胶电泳检测提取的DNA浓度和纯度,测定PCR产物灰度值。结果：三种方法都可以提取出基因组DNA,测定DNA浓度分别为：(35.56±15.27)ng/μL,(45.13±16.54)ng/μL,(57.93±14.5)ng/μL,组间存在统计学差异(P<0.01),A260/A280值分别为：1.46±0.19,1.49±0.16,1.519±0.23,三组间比较差异无统计学意义(P>0.05)；三种方法提取的基因组DNA都能通过PCR扩增出目的条带,PCR产物电泳灰度值结果分别为：75.421±3.435,149.32±6.875和229.52±19.55,组间比较差异有统计学意义(P<0.01)。结论：三种不同方法都能提取出冻存时间较长的非抗凝血块基因组DNA,但Invitrigen purelinkTM试剂盒法效果最好。