Array painting: a protocol for the rapid analysis of aberrant chromosomes using DNA microarrays.
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.
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