运用Identifiler试剂盒对硅珠法提取石蜡包埋组织DNA质量的评估
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江苏省教育厅科学基金资助(05KJD310140)


Using Identifiler Kit evaluates silica extracting DNA from paraffin-embedded tissue
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    目的:运用硅珠法从石蜡包埋的组织中提出DNA,Identifiler试剂盒评估DNA质量-方法:南京医科大学第一附属医院病理科2009~2011年石蜡包块49例,运用硅珠法提取石蜡包埋组织DNA,通过紫外分光光度计测定DNA纯度和Identifiler试剂盒PCR后3130型毛细管电泳仪分析提取DNA片段的完整性-结果:经紫外分光光度计测定胃癌组织D(260 nm)/D(280 nm)均值为1.84 ± 0.02,肺癌组织为1.85 ± 0.02,甲状腺癌组织为1.82 ± 0.02,3组间比较P > 0.05,因此硅珠法提取石蜡包埋组织DNA不会因组织差异而影响提取效果;Identifiler试剂盒PCR后毛细管电泳显示其提取效果,46例(93%)得出完整的16个基因座STR峰型-结论:硅珠法可应用于多种组织的DNA提取,并保证提取DNA样品的纯度和完整度-

    Abstract:

    Objective:To extract DNA from paraffin-embedded tissue by silica-extraction. Methods:The DNA samples were obtained via silica-extraction method from 49 cases of paraffin-embedded tumor tissues from Department of Pathology in the First Affiliated Hospital of NJMU. The 16 STR gene loci of PCR-DNA fragments were identified with Identifiler Kit. The purity of DNA extracted was analyzed by ultraviolet spectrophotometer. Results:The mean D(260 nm)/D(280 nm) of DNA extracted from gastric carcinoma was 1.84 ± 0.02,from pulmonary carcinoma is 1.85 ± 0.02,from thyroid carcinoma was 1.82 ± 0.02. There was no significant difference in D(260 nm)/D(280 nm) among three carcinoma type groups(P > 0.05). The 46/49 cases(93%) showed the full peaks of all the 16 STR gene loci of PCR-DNA fragment. Conclusion:Silica-extraction may be used to various kinds of paraffin-embedded tissues and extract DNA with high purity and high integrity.

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林 旭,王建文,尹雅莎,谢素梅,冀 强.运用Identifiler试剂盒对硅珠法提取石蜡包埋组织DNA质量的评估[J].南京医科大学学报(自然科学版),2012,(4):577-581

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  • 收稿日期:2011-11-29
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