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

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。

Separation of DNA Fragments with a Broad Range of Molecular Weight by Capillary Electrophoresis with Sieving Matrix of Poly(ethylene oxide)

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).