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利用单链构象多态性(SSCP),建立微流控芯片电泳(ME)联合激光诱导荧光检测(LIF)技术,检测人类p53基因点突变的方法。设计不同碱基长度的p53单链序列,针对易突变的外显子7,8,9进行SSCP分析,分离野生与突变的单链DNA序列;研究了筛分介质聚乙烯基氧化物(PEO)的浓度,场强对芯片电泳行为的影响。在PEO质量分数为0.5%,分离场强为260V/cm时,100 s之内就可以实现样品p53外显子7,8,9的野生型与突变型碱基的分离检测。
Single-strand conformation polymorphism (SSCP) was used to establish a method of detecting human p53 gene mutation by microfluidic chip electrophoresis (ME) combined with laser-induced fluorescence detection (LIF). The single-stranded DNA sequence of p53 with different base length was designed. SSCP analysis was performed on the exon 7,8,9 which was easy to mutate, and the wild-type and single-stranded DNA sequences were isolated. The effect of screening medium PEO, The concentration of field strength on the chip electrophoresis behavior. When the mass fraction of PEO was 0.5% and the separation field intensity was 260 V / cm, the separation of wild-type and mutant bases of p53 exons 7, 8 and 9 was detected within 100 s.