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目的研究运用改良酚-氯仿法从甲醛固定组织中提取DNA的方法,对比心、肝、肺、脑4种脏器DNA提取质量的优劣,从而了解哪种经甲醛固定后的组织易于得到质量较为可靠的DNA,以达到对DNA扩增及后续研究的目的。方法选取甲醛固定标本脏器各14例,以蛋白酶K消化和酚-氯仿法提取检材中DNA,对所得DNA质量以紫外分光光度计、PCR扩增及琼脂糖凝胶电泳分析进行结果判断。结果心、肝、肺、脑4种组织所提DNA的OD_(260)/OD_(280)比值分别为(1.842±0.380)、(1.861±0.076)、(1.387±0.113)、(1.428±0.087)。每100毫克组织中DNA含量(μg)分别为(0.944±0.530)、(1.096±0.544)、(0.348±0.273)、(0.601±0.238)。PCR扩增后经琼脂糖凝胶电泳分析显示,心肌和肝脏组织的条带清晰度高于肺脏和脑组织。结论经甲醛浸泡固定的心肌组织和肝脏组织,运用改良酚-氯仿法所得DNA质量较为可靠,可以用于后续研究。
Objective To study the method of DNA extraction from formalin-fixed tissues by modified phenol-chloroform method, and to compare the quality of DNA extracted from four kinds of organs including heart, liver, lung and brain, so as to know which kind of tissues are easy to obtain after being fixed by formaldehyde More reliable DNA, in order to achieve the purpose of DNA amplification and follow-up study. Methods Forty fourteen cases of formaldehyde fixed specimens were selected. DNA was extracted by digestion with proteinase K and phenol - chloroform. The quality of the obtained DNA was judged by ultraviolet spectrophotometer, PCR amplification and agarose gel electrophoresis. Results The OD_ (260) / OD 280 ratios of DNA extracted from the four tissues of heart, liver, lung and brain were (1.842 ± 0.380), (1.861 ± 0.076), (1.387 ± 0.113) and (1.428 ± 0.087) . The DNA content (μg) in each 100 mg tissue was (0.944 ± 0.530), (1.096 ± 0.544), (0.348 ± 0.273) and (0.601 ± 0.238), respectively. After PCR amplification, agarose gel electrophoresis analysis showed that the clarity of the bands in the myocardium and liver tissues was higher than that in the lungs and brain tissues. Conclusion The DNA quality obtained by modified phenol - chloroform method is more reliable after being soaked in formaldehyde for myocardial tissue and liver tissue, and can be used for subsequent study.