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Doctoral Thesis
DOI
10.11606/T.9.2017.tde-19052017-121409
Document
Author
Full name
Evelyn Anzai Kanto
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2007
Supervisor
Committee
Hirata, Mario Hiroyuki (President)
Cicarelli, Regina Maria Barreto
Gattas, Gilka Jorge Figaro
Nunes, Fabio Daumas
Oliveira, Rogerio Nogueira de
Title in Portuguese
Caracterização e desenvolvimento de sistemas de referências alélicas de loci de STR para controle de qualidade em identificação humana
Keywords in Portuguese
Biologia molecular (Aplicações)
Biomarcadores (Controle de qualidade)
Controle de Qualidade
DNA (Aplicações; Controle de qualidade)
Identificação Humana
Marcador Molecular
STR
Abstract in Portuguese
Reprodutibilidade, sensibilidade e controle de qualidade são essenciais em identificação humana pelos ácidos nucléicos exigindo uma referência que seja estável a temperatura ambiente e que monitore todo procedimento de manipulação da amostra. O estudo se propõe a construção de referências alélicas com plasmídeos recombinantes de 13 loci de STRs recomendados pelo CODIS-CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51 e D21S11. Os ácidos nucléicos foram extraídos de amostras de sangue ou saliva, com caracterização das STRs pela PCR e análise de fragmento. Os fragmentos dos diferentes alelos foram clonados em plasmídeos pGEM-T. Estes clones foram avaliados, utilizando-se dois sistemas comerciais. Obteve-se amplificações dos alelos das STRs D3S1358, TH01, D21S11, D5S818, D13S317 e VWA pelo PowerPlex®16 e D18S51, D21S11, D3S1358, D5S8181, D8S1179, TH01 e TPOX pelo AmpFLSTR®IdentifilerTM. A referência alélica com plasmídeos recombinantes foi construída e devidamente caracterizada demonstrando o seu potencial de aplicação como controle positivo.
Title in English
Caracterization and development of STRs allelic reference system to quality control in human identification
Keywords in English
Biomarcadores (Quality control)
Control
DNA (Applications; Quality control)
Human identification
Molecular biology (Applications)
Molecular marker
STR
Abstract in English
Reproducibility, sensibility and quality control are essentials in DNA human identification, requiring a stabile ambient temperature reference that monitors all procedures of samples manipulation. This study propose allelic reference construction, that will be useful as externai positive control for 13 CODIS STRs - CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, e D21S11. The nucleic acid was extracted from saliva or blood samples and enabling PCR-based typing and fragment analysis. Differents alleles founded were cloned into pGEM-T easy vector plasmids. The clones were analyzed using two commercial systems. The STRs D3S1358, TH01, D21S11, D5S818, D13S317 and VWA were amplified by PowerPlex®16, and D18S51, D21S11, D3S1358, D5S8181, D8S1179, TH01 and TPOX by AmpFLSTR®IdentifilerTM. The allelic reference with recombinants plasmids was constructed and. duly characterized, and the positive control potential application was demonstrated.
 
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Publishing Date
2017-05-19
 
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