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Infrared fluorescent automated detection of thirteen short tandem repeat polymorphisms and one gender‐determining system of the CODIS core system
Author(s) -
Ricci Ugo,
Sani Ilaria,
Guarducci Silvia,
Biondi Cristina,
Pelagatti Sara,
Lazzerini Valentina,
Brusaferri Alessandra,
Lapini Manuela,
Andreucci Elena,
Giunti Laura,
Uzielli Maria Luisa Giovannucci
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200011)21:17<3564::aid-elps3564>3.0.co;2-o
Subject(s) - genetics , microsatellite , multiplex , primer (cosmetics) , dna sequencer , biology , typing , polymerase chain reaction , multiplex polymerase chain reaction , population , computational biology , dna profiling , str analysis , dna , allele , microbiology and biotechnology , gene , chemistry , medicine , environmental health , organic chemistry
We used an infrared (IR) automated fluorescence monolaser sequencer for the analysis of 13 autosomal short tandem repeat (STR) systems (TPOX, D3S1358, FGA, CSF1PO, D5S818, D7S820, D8S1179, TH01, vWA, D13S317, D16S359, D18S51, D21S11) and the X‐Y homologous gene amelogenin system. These two systems represent the core of the combined DNA index systems (CODIS). Four independent multiplex reactions, based on the polymerase chain reaction (PCR) technique and on the direct labeling of the forward primer of every primer pair, with a new molecule (IRDye TM 800), were set up, permitting the exact characterization of the alleles by comparison with ladders of specific sequenced alleles. This is the first report of the whole analysis of the STRs of the CODIS core using an IR automated DNA sequencer. The protocol was used to solve paternity/maternity tests and for population studies. The electrophoretic system also proved useful for the correct typing of those loci differing in size by only 2 bp. A sensibility study demonstrated that the test can detect an average of 10 pg of undegraded human DNA. We also performed a preliminary study analyzing some forensic samples and mixed stains, which suggested the usefulness of using this analytical system for human identification as well as for forensic purposes.