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Analysis of multiplexed short tandem repeat (STR) systems using capillary array electrophoresis
Author(s) -
Mansfield Elaine S.,
Robertson James M.,
Vainer Marina,
Isenberg Alice R.,
Frazier Rachael R.,
Ferguson Karin,
Chow ShiTse,
Harris Dennis W.,
Barker David L.,
Gill Peter D.,
Budowle Bruce,
McCord Bruce R.
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.1150190118
Subject(s) - capillary electrophoresis , genotyping , multiplex , str analysis , str multiplex system , multiplex polymerase chain reaction , microsatellite , polymerase chain reaction , dna sequencer , chromatography , genotype , microbiology and biotechnology , biology , analytical chemistry (journal) , genetics , allele , chemistry , gene
The profiling of polymorphic short tandem repeat (STR) markers is being applied to human identification, parentage testing and genetic mapping. Reliable genotyping of these markers is facilitated by polymerase chain reaction (PCR) amplification and high‐resolution electrophoretic separation. Capillary array electrophoresis (CAE) offers very rapid, high‐resolution separation of the amplified DNA and potential for automated sample processing not realized employing conventional slab‐gel electrophoresis. The use of CAE to type DNA samples amplified at 11 genetic loci in multiplex profiles is presented. Two sets totaling 208 samples were amplified in a multiplex fashion using Amp Fl STR‐Blue or Amp Fl STR‐Green I and analyzed in a blind study using CAE. With the exception of one sample, the CAE genotyping results were in complete agreement with results obtained using a single‐capillary system or two slab‐gel electrophoresis systems. The sample, genotype TH01 7/10, migrated similar to TH01 6.3/9.3 allele sizes, which suggested a potential band migration shift. The recommended approach to such an observation is to analyze the sample again. The sample was rerun and correct genotype verified. Allelic ladder samples were analyzed multiple times by CAE to determine sizing accuracy and precision. The sizing of over 240 allelic ladder samples vielded an average within‐run precision of ± 0.13 bp and between‐run precision of ± 0.21 bp for fragments up to 350 bp. The CAE protocols permit processing of up to 96 multiplex STR samples in under 70 min.