Capillary sample introduction of polymerase chain reaction (PCR) products separated in ultrathin slab gels

被引:11
作者
Bullard, KM [1 ]
Hietpas, PB [1 ]
Ewing, AG [1 ]
机构
[1] Penn State Univ, Dept Chem, University Pk, PA 16802 USA
关键词
capillary electrophoresis; ultrathin slab gels; polymerase chain reaction products; parallel separations; forensic analysis;
D O I
10.1002/elps.1150190113
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 mu m 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 separtion resolution still needs to be improved.
引用
收藏
页码:71 / 75
页数:5
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