聚丙烯酰胺凝胶DNA水平电泳检测多药耐药基因多态性

Detection of multi-drug resistant gene1 single nucleotide polymorphisms with DNA electrophoresis on horizontal polyacrylamide gels

目的探讨聚丙烯酰胺凝胶(PAGE)DNA水平电泳条件,分析多药耐药基因单核苷酸多态性(MDR1SNPs)。方法以DNA标准为标本探讨PAGE DNA水平电泳条件;PCR-限制性片段长度多态性作为健康人MDR1SNPs C3435T分析,PAGE DNA水平电泳作PCR产物酶切结果分析。结果10 mL PAGE含40%过硫酸铵70μL,四甲基乙二胺20μL,隔绝空气在琼脂糖水平电泳槽制胶。交联度为29:1,凝胶浓度为7.5%时,效果最好。45例健康人MDR1C3435T中,20例为C/C型、T/C型17例、T/T型8例。结论PAGE DNA水平电泳简便易行,分离效果好,可用于MDR1SNPs分析,适合基层推广。

Objective To investigate the condition of DNA electrophoresis on horizontal polyaceylamide gels （PAGE） and detect multi drug resistant genel single nucleotide polymorphisms （MDR1SNPs） with the method. Methods To investigate the condition of PAGE DNA horizontal elec trophoresis with DNA markers; analysis MDR1SNPs C3435T of Han population with PCR-restriction fragment length polymorphisms（PCR-RFLP） and detect the fragments length of the PCR products di gested by MboI. Results 10 mL of PAGE contains 70 μL of 40% Ammonium Persuifate and 20 μL of TEMED and prepared the gel in an agarose horizontal electrophoresis tray without air. The efficiency is the best with the gel combination rate 29： t and the concentration of 7. 5%. The gene types of MDR1SNPs C3435T in 45 healthful people are： type C/C, 20;T/C,i7;T/T,8. Conclusion It is sire ple,and efficient to detect gene types of MDR1SNPs with PAGE DNA horizontal electrophoresis. It could be recommended to use in laboratories lack of resource.