基于玻璃电泳芯片分离STR片段的研究.pdfVIP

中国科技论文在线 基于玻璃电泳芯片分离 STR 片段的研究 艾洪新，何越，荣吉赞，杨楠** （凯晶生物科技（苏州）有限公司，江苏 苏州 215123） 5 摘要：在普通玻璃材料的电泳芯片上，利用激光诱导荧光检测系统实现了对单链 STR （短 串联重复序列）片段快速高效的分离检测和优化研究。选用分离通道为8.5cm 玻璃芯片，用 ABI 公司生产的 POP-4 和 POP-7 筛分胶来分离法医鉴定试剂盒扩增的DNA 片段。通过改变 酸碱清洗芯片的顺序研究清洗芯片对于分离 STR 片段的影响，通过改变电压及筛分胶的配 比来优化筛分效果，提高分离度。实验结果表明，改变酸碱清洗顺序影响出峰时间和分离度， 10 将 POP-4 和 POP-7 混合使用比单独使用效果要好，改变电压对分离度也有一定影响。 关键词：电泳芯片；激光诱导荧光检测系统；STR 片段；酸碱清洗；POP-4 和 POP-7 中图分类号：O657 Electrophoretic Analysis of Short Tandem Repeats (STR) in 15 glass chip Aihongxin, Heyue, Rongjizan, Yangnan (ChemoGene Biotechnologies Suzhou Ltd, JiangSu SuZhou 215123) Abstract: The aim of the study is to investigate the factors that affect separation efficiency of STR single-stranded DNA fragments on glass chip-capillary electrophoresis (chip-CE), detected by 20 single-colour laser-induced fluorescence detection system. The STR fragments were amplified by AmpFlSTR® Identifiler® Plus Kit (ABI) in a common PCR thermal cycler, and the fragments were analyzed through POP-4 and POP-7 gel-matrix mixture (ABI) in a glass channel (8.5 cm long) on a glass chip. The experiments showed that the order of glass channel acid and alkali cleaning procedure has greater impact on the retention time and resolution; mixture of POP-4 and 25 POP-7 obtained better resolution than that of using POP-4 or POP-7 alone; and separation voltage also influences the separation resolution. Keywords: chip-CE; laser-induced fluorescence detec