RESTRICTION MAPPING OF GENES BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION

Restriction mapping is one of the essential steps in gene analysis and molecular biology studies, Slab gel electrophoresis is the traditiona l way to separate DNA fragments for restriction mapping, However, slab gel electrophoresis does not provide sufficient resolution as require d in many mapping applications, and the use of radioisotopes in tradit ional mapping methods creates health hazards, In the present study, ca pillary electrophoresis coupled with laser-induced fluorescence detect ion and a modified partial digestion mapping procedure was developed t o map DNA fragments. By using capillary electrophoresis, a restriction map of a genomic lambda phage clone of human interleukin 5 receptor a chain (IL5R alpha) gene was constructed, The IL5R alpha gene was anal yzed to have five XbaI enzyme cutting sites at locations 1370, 2290, 2 950, 5430, and 9330, The system was further characterized by using pBl uescript SK(+) phagemid DNA as a model, Using a sequence-derived map a s a reference, the pBluescript SK(+) restriction map constructed by ca pillary electrophoresis had an accuracy greater than 90%.