| 包晗赵锦荣汪钦郭晏海刘永兰雷小英梁平孙建斌颜真△.一种简易SNP 检测方法的建立[J].,2011,11(10):1969-1971 |
| 一种简易SNP 检测方法的建立 |
| A Simplified Method of SNP Analysis |
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| DOI： |
| 中文关键词: 硝酸纤维素膜 核酸扩增 SNP 生物素- 亲和素酶联显色 |
| 英文关键词: Nitrocellulose membrane Nucleic acid amplification SNP ABS-ELISA |
| 基金项目:国家“十一五”重大传染病专项(2009ZX10004-311)；国家重点基础研究发展规划（973）项目(2010CB933900);
西安市科技计划(SF09027) |
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| 中文摘要: |
| 目的：建立一种快速、简单的SNP（Single Nucleotide Polymorphisms）检测方法。方法：设计带生物素标记的扩增引物对检测
用具有单碱基差异的野生型和突变型靶序列分别进行扩增，然后通过紫外交联的方式将相应检测靶序列的探针固定在硝酸纤维
素膜上，借助Taq 酶完成膜上单引物延伸，从而对探针捕获的靶序列进行延伸固定在膜上，最后使用生物素- 亲和素酶联显色
(ABS-ELISA)反应肉眼观察结果。结果：阳性和阴性对照探针显示正常。野生型探针和突变型探针能够分别特异性结合靶序列，并
通过生物素和亲和素显色系统放大为一种肉眼可判断结果的检测形式。结论：建立了一种基于硝酸纤维素膜载体上进行核酸扩
增的SNP 检测方法。 |
| 英文摘要: |
| Objective: To build a fast and simple method of Single Nucleotide Polymorphisms (SNP) analysis. Methods: The
wild-type and mutant DNA sequences were amplified with single base pair mismatch by biotin labeled primers. Probes were crosslinked
onto nitrocellulose membrane by UV irradiation. Complementary target with biotin labeled sequence can be extend and fixed after
combined with probes on membrane by Taq DNA polymerase enzyme. After ABS-ELISA coloration, results were observed directly.
Results: Normal working system was showed by the negative and positive blots. Wild-type and mutant probe can distinguish single base
pair mismatch. And the coloration result can be judged directly. Conclusion: A method for detecting SNPs with nucleic acid
amplification based on nitrocellulose membrane was established. |
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