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Capillary sample introduction of polymerase chain reaction (PCR) products separated in ultrathin slab gels
Author(s) -
Bullard Katherine M.,
Hietpas Paula Beyer,
Ewing Andrew G.
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150190113
Subject(s) - ethidium bromide , polymerase chain reaction , reproducibility , locus (genetics) , capillary electrophoresis , chromatography , microbiology and biotechnology , sample preparation , analytical chemistry (journal) , chemistry , biology , dna , genetics , gene
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 μm 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 42s apart for each of the three consecutive separations. This technique shows potential to increase sample throughput in STR techniques although separation resolution still needs to be improved.