Day I N, Spanakis E, Chen X, O'Dell S D
Wessex Human Genetics Institute, Southampton University Hospitals NHS Trust, UK.
Electrophoresis. 1999 Jun;20(6):1250-7. doi: 10.1002/(SICI)1522-2683(19990101)20:6<1250::AID-ELPS1250>3.0.CO;2-5.
Molecular genetic epidemiology, association and linkage studies in populations, human or other species, is now yielding powerful new insights into disease and susceptibility genes. Inter alia, the subject requires laboratory analytical methodologies focused on achieving high throughput. Here we review one suite of methodology suitable for such laboratories. Microplate array diagonal gel electrophoresis (MADGE) was invented for molecular genetic epidemiological studies. It combines direct compatibility with microplates, convenient polyacrylamide gel electrophoresis and economy of time and reagents, at minimal capital cost, and enables one user to run up to several thousand gel lanes per day for direct assay of single-base variations. Melt-MADGE combines temporal thermal ramp apparatus to achieve similar throughput for de novo mutation scanning.
分子遗传流行病学,以及在人类或其他物种群体中的关联和连锁研究,正在为疾病和易感基因带来强大的新见解。尤其重要的是,该领域需要专注于实现高通量的实验室分析方法。在此,我们综述一套适用于此类实验室的方法。微孔板阵列对角线凝胶电泳(MADGE)是为分子遗传流行病学研究而发明的。它结合了与微孔板的直接兼容性、便捷的聚丙烯酰胺凝胶电泳以及时间和试剂的经济性,成本最低,使一名用户每天能够运行多达数千个凝胶泳道,用于直接检测单碱基变异。熔解-MADGE结合了时间热梯度装置,以实现类似的从头突变扫描通量。