TP53 Sequencing and XAF1 Variant Analysis

Genomic DNA was isolated and used to determine TP53 sequence and the XAF1 p.E134∗ variant status as previously reported.⁸ Additionally, fluorescent-labelled PCR products for 10 polymorphic microsatellite markers located within (VNTRp53) or telomeric to the TP53 locus (Table S1) were genotyped by using the ABI 3500 sequencer (ThermoFisher Scientific) and sizes assigned to the different fragments using GeneMapper v 6.0 (ThermoFisher Scientific).⁸ ^,¹³ (link) Haplotypes were determined by segregation analysis of genomic DNA from parents and descendants, loss of heterozygosity in tumor samples, or inferred.

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Pinto E.M., Fridman C., Figueiredo B.C., Salvador H., Teixeira M.R., Pinto C., Pinheiro M., Kratz C.P., Lavarino C., Legal E.A., Le A., Kelly G., Koeppe E., Stoffel E.M., Breen K., Hahner S., Heinze B., Techavichit P., Krause A., Ogata T., Fujisawa Y., Walsh M.F., Rana H.Q., Maxwell K.N., Garber J.E., Rodriguez-Galindo C., Ribeiro R.C, & Zambetti G.P. (2023). Multiple TP53 p.R337H haplotypes and implications for tumor susceptibility. Human Genetics and Genomics Advances, 5(1), 100244.

Publication 2023

ABI 3500 sequencer GeneMapper v 6.0

Genomic Haplotypes Loss of heterozygosity Microsatellite markers Parents Polymorphic Telomeric Tp53 Tumor Xaf1

Corresponding Organization :

Other organizations : Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, Instituto de Tecnologia do Paraná, Hospital Sant Joan de Déu Barcelona, Cancer Genetics (United States), Research Network (United States), Instituto Português de Oncologia Francisco Gentil, IPO Porto, Universidade do Porto, Medizinische Hochschule Hannover, University of Pennsylvania, University of Michigan–Ann Arbor, Memorial Sloan Kettering Cancer Center, Universitätsklinikum Würzburg, Chulalongkorn University, National Health Laboratory Service, University of the Witwatersrand, Hamamatsu University School of Medicine

Top 5 similar protocols

Variable analysis

independent variables

TP53 sequence
XAF1 p.E134* variant status

dependent variables

Genotypes of 10 polymorphic microsatellite markers located within (VNTRp53) or telomeric to the TP53 locus

control variables

Genomic DNA from parents and descendants
Tumor samples

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