基于国产遗传分析仪的毛细管电泳筛分介质的制备

Study on the preparation method of capillary electrophoresis sieving medium based on domestic genetic analyzer

目的制备一种毛细管电泳筛分介质,将其应用于GA118-16A型遗传分析仪上。方法通过聚合反应、冷冻干燥得到白色固体聚丙烯酰胺(LPA),经溶胶缓冲液溶胀后获得所需的毛细管电泳筛分介质。对其分子量、结构等关键性能指标进行表征;并将其应用于GA118-16A型遗传分析仪上,根据空间校正、光谱校正及STR检测结果对其筛分性能进行评价。结果制备的筛分介质Mw 1.8×105Da,Mn 1.2×105Da,多分散性1.5;结构正确且纯度高;在GA118-16A型遗传分析仪上能够建立空间校正、光谱校正文件;能够有效分离相差1bp的DNA片段,峰型尖锐、无杂峰、无拖尾、无丢峰现象。结论该方法制备的筛分介质完全可应用于GA118-16A等国产遗传分析仪。

Objective To prepare a capillary electrophoresis sieving medium and apply it in GA118-16 A genetic analyzer.Methods The white solid polyacrylamide（LPA） was prepared by polymerization and lyophilized.Through the swelling of the sol buffer,the sieving medium was obtained.The sieving medium was evaluated by 1） characterizing the parameters,including molecular weight,structure and viscosity,2） applying in the GA118-16 A genetic analyzer,including the spatial calibration,the spectral calibration and the STR analysis..Results The prepared sieving medium Mw 1.8 x 105 Da,Mn 1.2 x 105 Da,is of correct structure and high purity.The polydispersity was 1.5 The spatial calibration and spectral calibration files can be established successfully in GA118-16 A genetic analyzer,and the sieving medium can effectively separate the DNA fragments with 1 bp difference.The STR profile is of sharp peaks,no impurity peaks,no tail,and no peak loss.Conclusion The sieving medium prepared by the method can be applied to domestic genetic analyzer such as GA118-16 A.