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 i.d.,有效长度21.0 cm)分离DL5000 DNA Marker(DNA长度为100～5000 bp),考察了筛分介质浓度、缓冲液pH值、分离电压和溴化乙锭浓度对分离双链DNA片段的影响,优化得到分离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5 mg/mL,缓冲液pH值8.0,电压-12.0 kV及溴化乙锭浓度3.0 μg/mL.在此条件下,可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)扩增产物同时进行检测,分离和鉴定效果良好.%DL5000 DNA marker fragments( 100-5000 bp) were separated by non-gel sieve capillary electrophoresis on a silanized fused silica capillary column(31.2 cm ×75 μm i.d. with 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 double-strand DNA fragments were investigated. The optimum conditions for separation of 100-5000 bp DNA fragments were established as 5 mg/mL poly ( ethylene oxide), pH = 8.0, 3.0 μg/mL ethidium bromide and voltage of - 12.0 kV. Under these conditions, the multiplex polymerase chain reaction (PCR) magnified products from the sorbitol dehydrogenase gene (SDH) and the ethylene receptor gene (ETR1) were simultaneously detected, and good identification and resolution were obtained.
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