Capillary sample introduction of polymerase chain reaction (PCR) products separated in ultrathin slab gels.
Polymerase chain reaction (PCR) amplified short tandem repeat (STR) samples from the HUMVWF locus have been analyzed using a unique sample introduction and separation technique. A single capillary is used to transfer samples onto an ultrathin slab gel (57 microm thin). This ultrathin nondenaturing polyacrylamide gel is used to separate the amplified fragments, and laser-induced fluorescence with ethidium bromide is used for detection. The feasibility of performing STR analysis using this system has been investigated by examining the reproducibility for repeated samples. Reproducibility is examined by comparing the migration of the 14 and 17 HUMVWF alleles on three consecutive separations on the 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 throughput in STR analysis techniques although separation resolution still needs to be improved.
Bullard
K M
KM
Department of Chemistry, The Pennsylvania State University, University Park 16802, USA.
Hietpas
P B
PB
Ewing
A G
AG
D020106:Acrylamide; D000178:Acrylamides; D004247:DNA / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005782:Gels; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D015203:Reproducibility of Results; D014841:von Willebrand Factor / Q000235:genetics
D000178:Acrylamides; D005782:Gels; D014841:von Willebrand Factor; D020106:Acrylamide; D004247:DNA