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Array painting: a protocol for the rapid analysis of aberrant chromosomes using DNA microarrays.
Gribble, Susan M; Ng, Bee Ling; Prigmore, Elena; Fitzgerald, Tomas; Carter, Nigel P.
Afiliação
  • Gribble SM; Human Genetics, Sulston Laboratories, The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridge, UK. smg@sanger.ac.uk
Nat Protoc ; 4(12): 1722-36, 2009.
Article em En | MEDLINE | ID: mdl-19893508
Array painting is a technique that uses microarray technology to rapidly map chromosome translocation breakpoints. Previous methods to map translocation breakpoints have used fluorescence in situ hybridization (FISH) and have consequently been labor-intensive, time-consuming and restricted to the low breakpoint resolution imposed by the use of metaphase chromosomes. Array painting combines the isolation of derivative chromosomes (chromosomes with translocations) and high-resolution microarray analysis to refine the genomic location of translocation breakpoints in a single experiment. In this protocol, we describe array painting by isolation of derivative chromosomes using a MoFlo flow sorter, amplification of these derivatives using whole-genome amplification and hybridization onto commercially available oligonucleotide microarrays. Although the sorting of derivative chromosomes is a specialized procedure requiring sophisticated equipment, the amplification, labeling and hybridization of DNA is straightforward, robust and can be completed within 1 week. The protocol described produces good quality data; however, array painting is equally achievable using any combination of the available alternative methodologies for chromosome isolation, amplification and hybridization.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Translocação Genética / Mapeamento Cromossômico Limite: Humans Idioma: En Revista: Nat Protoc Ano de publicação: 2009 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Translocação Genética / Mapeamento Cromossômico Limite: Humans Idioma: En Revista: Nat Protoc Ano de publicação: 2009 Tipo de documento: Article