作　　者： ; ; ; ; ; ; ; ; ;

机构地区： 兰州军区兰州总医院

出　　处： 《分析测试学报》 2009年第4期379-384,共6页

摘　　要： 建立了一种毛细管电泳快速高效检测聚合酶链反应(PCR)扩增产物以及限制性内切酶酶切产物的方法,使其更好地用于基因诊断。以聚环氧乙烷(poly(ethylene oxide),PEO)为筛分介质,用涂层的毛细管柱(37 cm×75μm,有效长度27 cm)分离pUC19 DNA/MspⅠ(HpaⅡ)Marker标准DNA片段。考察了筛分介质的质量浓度、pH值、毛细管柱的温度和运行电压。在1×TBE(pH8.2)电泳液、电压15 kV、温度15℃,于10 min内成功分离了Marker标准DNA片段。该方法快速、灵敏、准确,用于临床76例肺癌患者正常组织和肿瘤组织p53基因和ras基因点突变情况的检测,结果满意。 A rapid and high efficiency capillary electrophoresis method was developed for detecting the products of polymerase chain reaction （PCR） amplification and restriction enzyme digestion to preferably use for gene diagnosis. The pUC19 DNA/Msp Ⅰ （ Hpa U ） Marker standard DNA fragments were separated on a fused silica capillary column （37 cm × 75 μm, the effective length 27 cm） using poly（ ethylene oxide） （PEO） as sieving matrix. The influencing factors, including the concentration of buffer, pH value, the cartridge temperature and the applied potential were investigated. The best separation efficiency of the Marker standard DNA fragments was achieved in 10 min under using a running buffer of 1 × TBE（pH 8.2）, applied voltage of 15 kV and temperature of 15 ℃. The pro- posed method was rapid, accurate and convenient, and was successfully applied in the detection of the p53 and ras gene mutation of normal tissues and lung cancer tissues in 76 lung cancer patients.

关 键 词： 毛细管电泳 基因突变 肺癌 聚环氧乙烷 纯化

领　　域： [理学] [理学] [生物学]