CAPILLARY SAMPLE INTRODUCTION OF POLYMERASE CHAIN-REACTION (PCR) PRODUCTS SEPARATED IN ULTRATHIN SLAB GELS

Polymerase chain reaction (PCR) amplified short tandem repeat (STR) sa mples from the HUMVWF locus have been analyzed using a unique sample i ntroduction and separation technique. A single capillary is used to tr ansfer samples onto an ultrathin slab gel (57 mu m thin). This ultrath in nondenaturing polyacrylamide gel is used to separate the amplified fragments, and laser-induced fluorescence with ethidium bromide is use d for detection. The feasibility of performing STR analysis using this system has been investigated by examining the reproducibility for rep eated samples. Reproducibility is examined by comparing the migration of the 14 and 17 HUMVWF alleles on three consecutive separations on th e ultrathin slab gel. Using one locus, separations match in migration time with the two alleles 42 s apart for each of the three consecutive separations. This technique shows potential to increase sample throug hput in STR analysis techniques although separtion resolution still ne eds to be improved.