ANALYSIS OF SICKLE-CELL GENE USING POLYMERASE CHAIN-REACTION AND RESTRICTION ENZYME BSU-361

A 772bp DNA fragment from human beta-globin gene has been amplified hy polymerase chain reaction (PCR) and subjected to restriction enzyme a nalysis using Bsu 361, an isoschizomer of restriction enzyme Mst II. T his protocol has been designed basically to enhance the analytical fac ility. for the detection of sickle cell mutation. A 430bp DNA fragment was found to be associated with the mutant locus, whereas 228bp and 2 02bp DNA fragments were generated from the normal locus, This differen ce of about 202bp in the resulting fragments from the mutant and norma l loci has improved discriminatory power in the genotype analysis of t he sickle cell mutation.