不同粪便DNA提取试剂盒及样本储存时间对粪便人基因组DNA提取率的影响

目的比较不同粪便DNA提取试剂盒及粪便样本不同储存时间对粪便人基因组DNA提取效率的影响。方法对18例体检健康者使用QIAamp DNA Stool Mini Kit,MO BIO UltraCleanFecal DNA Isolation Kit,E.Z.N.A.Mag Bind Stool DNA Kit 3种粪便DNA提取试剂盒进行粪便DNA提取,通过紫外分光光度法对总DNA产量和纯度进行分析,通过实时荧光聚合酶链反应(PCR)方法对提取产物中结肠腺瘤性息肉病基因进行定量,并比较人基因组DNA提取效率的差异及不同样本储存时间对其影响。结果在3种试剂盒中,QIAamp DNA Stool Mini Kit对人基因组DNA的提取产量最高(平均67.81 ng/g),E.Z.N.A.Mag Bind Stool DNA Kit提取产物纯度最高(A260/A280平均2.07),MO BIO UltraCleanFecal DNA Isolation Kit操作耗时最短(约40 min);粪便样本在-70℃条件下储存30 d内,不同储存时间对总DNA提取效率差异有统计学意义(P0.01)。结论 QIAamp DNA Stool Mini Kit对粪便人基因组DNA的提取效率最高,且于-70℃储存粪便样本30 d内对人基因组DNA提取效率无影响。

Influence of Different Storage Times and DNAzol Reagent Kits on the Efficiency of Human Genomic DNA Extraction from Stools

Objective To compare the influence of different DNAzol reagent kits and storage times on the efficiency of hu- man genomic DNA extraction from stools. Methods DNA was extracted from stools of 18 healthy people following medical examina- tions by using three different stool DNA extraction kits （ QIAamp DNA Stool Mini Kit, MO BIO UltraClean Fecal DNA Isolation Kit and E. Z. N.A. Mag Bind Stool DNA Kit）. The quantity and purity of the total DNA were analyzed with ultraviolet spectrophotome- try, and the quantitation of the adenomatous polyposis colii （APC） gene in the total DNA was detected with real-time polymerase chain reaction （PCR） for comparing the difference of human genomic DNA extracting efficiency and the influence by different stor- age times. Results Among the three different kits, QIAGEN DNA Stool Mini kit showed the highest yield of human genomic DNA （averaged 67.81 ng/g） ; the E. Z. N.A. Mag Bind Stool DNA kit had the highest purity （A260/A280 averaged 2.07） ; MO BIO U1- traClean Fecal DNA Isolation kit had the shortest operative timesuck （about 40 min for a sample）. The stool samples were stored at - 70℃ within 30 d, the differences of efficiency of total DNA extraction were statistically significant （ P 〈 0. 01 ）, while the differ- ences of efficiency of human genomic DNA extraction were not statistically significant in terms of different storage times （ P 〉 0.05 ）. Conclusion QIAGEN kit has the highest efficiency for human genomic DNA extraction from stools, and stool samples store at -70℃ within 30 d show no influence on the extraction efficiency of human genomic DNA.