论文部分内容阅读
目的 :探讨提取大鼠毛发核酸的可靠方法。方法 :分别用PCR缓冲液法、SDS 蛋白酶K法和Chelex 10 0法从大鼠毛发中提取核酸 ,对所提核酸进行PCR扩增及电泳分析 ,并对 3种方法进行比较。结果 :从毛囊提取mtDNA的成功率 ,SDS 蛋白酶K法 (2 2 .5 0 % )分别与PCR缓冲液法 (6 4 .17% )和Chelex 10 0法(6 7.5 0 % )比较 ,差异均有统计学意义 (χ12 =4 2 .4 2 1,χ2 2 =2 8.80 0 ,均P <0 .0 1) ;PCR缓冲液法与Chelex 10 0法比较 ,差异无统计学意义 (χ2 =0 .2 96 ,P >0 .0 5 )。提取核DNA的成功率 ,SDS 蛋白酶K法 (6 0 .0 0 % )分别与PCR缓冲液法 (90 .0 0 % )和Chelex 10 0法 (90 .83% )比较 ,差异均有统计学意义 (χ12 =4 9.0 91,χ2 2 =30 .76 7,均P<0 .0 1) ;PCR缓冲液法与Chelex 10 0法比较 ,差异无统计学意义 (χ2 =0 .0 4 8,P >0 .0 5 )。PCR缓冲液法不能从毛干中提取核酸 ,SDS 蛋白酶K法不能从 1、2根鼠毛中提取mtDNA ,而Chelex 10 0法可从毛干和单根鼠毛中提取mtDNA和DNA。结论 :Chelex 10 0法对从毛发中提取核酸较为合适。
Objective: To explore a reliable method of extracting rat hair nucleic acid. Methods: Nucleic acid was extracted from rat hair by PCR buffer method, SDS protease K method and Chelex 10 0 method respectively. The nucleic acid was amplified by PCR and analyzed by electrophoresis. The three methods were compared. RESULTS: The success rate of extracting mtDNA from hair follicles was significantly lower than that of PCR buffer method (6.41%) and Chelex 10 method (65.5%) by SDS proteinase K assay (12.5% (Χ2 = 4 2 .4 2 1, χ2 2 = 2 8.80 0, all P <0.01). There was no significant difference between the PCR method and Chelex 10 0 method (χ2 = 0 .2 96, P> 0. 05). The success rate of nuclear DNA extraction, SDS protease K method (60.0%) were compared with the PCR buffer method (90.0%) and Chelex10 (90.83%), the differences were statistically Significance (χ 12 = 4 9.0 91, χ 2 2 = 30.776 7, all P <0.01); there was no significant difference between the PCR method and the Chelex 10 0 method (χ 2 = 0.408, P> 0 .0 5). PCR buffer method can not extract nucleic acids from the hair shaft, SDS protease K method can not extract mtDNA from 1, 2 mouse hair, and Chelex 10 0 method can be extracted from hairy and single rat hair mtDNA and DNA. Conclusion: The Chelex 10 0 method is more suitable for nucleic acid extraction from the hair.