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聚环氧乙烷无胶筛分毛细管电泳分离宽分子量范围DNA片段
引用本文:李玉荣,陈长宝,周杰. 聚环氧乙烷无胶筛分毛细管电泳分离宽分子量范围DNA片段[J]. 高等学校化学学报, 2011, 32(4): 844-850
作者姓名:李玉荣  陈长宝  周杰
作者单位:山东农业大学化学与材料科学学院, 泰安 271018
基金项目:国家自然科学基金(批准号:30871756)资助
摘    要:在无胶筛分毛细管电泳中,以聚环氧乙烷为筛分介质,用硅烷化处理的毛细管柱(31.2 cm×75 μm有效长度21.0 cm)分离DL5000 DNA Marker(DNA长度为100~5000 bp),研究筛分介质浓度、缓冲液pH、分离电压和溴化乙锭浓度对分离双链DNA片段的影响,优化出分离100~5000 bp DNA片段的最佳条件。毛细管电泳的最佳条件为PEO浓度0.5%、缓冲液pH值8.0、电压12 kV、溴化乙锭浓度3.0 μg/mL。此条件下,对山梨醇脱氢酶基因(SDH)和乙烯受体基因(ETR1)的聚合酶链式反应(PCR)扩增产物同时检测,分离、鉴定效果良好。

关 键 词:无胶筛分毛细管电泳  聚环氧乙烷  DNA分离  
收稿时间:2010-08-02

Separation of DNA Fragments with a Broad Range of Molecular Weight by Capillary Electrophoresis with Sieving Matrix of Poly(ethylene oxide)
LI Yu-Rong,CHEN Chang-Bao,ZHOU Jie. Separation of DNA Fragments with a Broad Range of Molecular Weight by Capillary Electrophoresis with Sieving Matrix of Poly(ethylene oxide)[J]. Chemical Research In Chinese Universities, 2011, 32(4): 844-850
Authors:LI Yu-Rong  CHEN Chang-Bao  ZHOU Jie
Affiliation:College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, China
Abstract:DL5000 DNA Marker (100~5000 bp) fragments were separated through no-gel sieve capillary electrophoresis on a fused silica capillary column (31.2 cm×75 μm, the effective length 21.0 cm) using Poly (ethylene oxide) as sieve matrix. The influences of Poly (ethylene oxide) concentration, pH value of running buffer, separation voltage and ethidium bromide concentration on the separation efficiency of different lengths of DNA fragments were investigated. The optimum conditions for 100~5000 bp DNA fragments were established as 0.5% Poly (ethylene oxide), pH 8.0, 3.0 μg/mL ethidium bromide and voltage of 12 kV. Under these conditions, good resolution was obtained in separating multiplex polymerase chain reaction (PCR) magnified products from sorbitol dehydrogenase gene (SDH) and ethylene receptor gene (ETR1).
Keywords:Non-gel sieve capillary electrophoresis  Poly(ethylene oxide)  DNA separation  
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