| 毛细管电泳-限制性片段长度多态性分析检测胃癌H-ras基因点突变 |
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| 引用本文： | 刘圆圆,王荣,贾正平,郭志强,辛晓婷,谢华,马骏,王娟,李文斌. 毛细管电泳-限制性片段长度多态性分析检测胃癌H-ras基因点突变[J]. 化学学报, 2009, 67(4): 323-328 |
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| 作者姓名： | 刘圆圆 王荣 贾正平 郭志强 辛晓婷 谢华 马骏 王娟 李文斌 |
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| 作者单位： | 兰州军区兰州总医院临床药理基地,兰州,730050;甘肃省红十字血液中心兰州730046;兰州大学生命科学学院,兰州,730000;兰州军区兰州总医院临床药理基地,兰州,730050;兰州军区兰州总医院临床药理基地,兰州,730050;兰州大学生命科学学院,兰州,730000 |
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| 基金项目： | 国家自然科学基金,甘肃省自然科学基金,中国博士后科学基金 |
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| 摘 要： | 建立一种毛细管电泳快速高效检测限制性内切酶酶切产物的方法, 使其更好地用于基因诊断. 以甲基纤维素(Methyl cellulose, MC)为筛分介质, 用pUC19 DNA/Msp I (Hpa II) Marker标准DNA片段为实验对象, 通过考察筛分介质的浓度、pH值、毛细管的温度和运行电压优化出分离小于600 bp的双链DNA片段的最适条件, 并将此方法应用于临床59例胃癌患者肿瘤组织H-ras基因12位密码子点突变情况的检测. MC是一种良好的筛分介质, 运用其进行毛细管电泳对于遗传性疾病的诊断将更加快速、准确、简便、灵敏.
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| 关 键 词： | 毛细管电泳 甲基纤维素 限制性片段长度多态性 H-ras 基因突变 |
| 收稿时间： | 2008-01-02 |
| 修稿时间： | 2008-08-27 |
H-ras Gene Mutation Detection of Gastric Cancer by Restriction Fragment Length Polymorphism with Capillary Electrophoresis |
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| Liu Yuanyuan,Wang Rong,Jia Zhengping,Guo Zhiqiang,Xin Xiaoting,Xie Hua,Ma Jun,Wang Juan,Li Wenbin. H-ras Gene Mutation Detection of Gastric Cancer by Restriction Fragment Length Polymorphism with Capillary Electrophoresis[J]. Acta Chimica Sinica, 2009, 67(4): 323-328 |
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| Authors: | Liu Yuanyuan Wang Rong Jia Zhengping Guo Zhiqiang Xin Xiaoting Xie Hua Ma Jun Wang Juan Li Wenbin |
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| Affiliation: | a Department of Pharmacy;Lanzhou General Hospital of PLA;Lanzhou 730050;b Gansu Red Cross Blood Center;Lanzhou 730046;c Life Science College;Lanzhou University;Lanzhou 730000 |
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| Abstract: | To establish the high performance detection method for separating products of restrictive enzyme DNA fragments and diagnosing mutation gene of gastric cancer by restriction fragment length polymorphism with capillary electrophoresis, methyl cellulose was used to separate pUC19 DNA/Msp I (Hpa II) fragments through capillary electrophoresis as sieving matrix. The effects of the concentration of methyl cellulose solution, running pH, running temperature and electric field strength on dsDNA (＜600 bp) analysis were investigated in the experiment. The optimum separation condition was obtained and applied to mutation detection of H-ras gene at codon 12 of 59 gastric cancer tissues. Methyl cellulose is an excellent medium and it will make diagnosis of genetic disease rapidly, exactly, conveniently and sensitively. |
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| Keywords: | H-ras |
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