Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes.
Nat Commun
; 6: 7147, 2015 May 12.
Article
em En
| MEDLINE
| ID: mdl-25962338
ABSTRACT
Fluorescence in situ hybridization (FISH) is a powerful single-cell technique for studying nuclear structure and organization. Here we report two advances in FISH-based imaging. We first describe the in situ visualization of single-copy regions of the genome using two single-molecule super-resolution methodologies. We then introduce a robust and reliable system that harnesses single-nucleotide polymorphisms (SNPs) to visually distinguish the maternal and paternal homologous chromosomes in mammalian and insect systems. Both of these new technologies are enabled by renewable, bioinformatically designed, oligonucleotide-based Oligopaint probes, which we augment with a strategy that uses secondary oligonucleotides (oligos) to produce and enhance fluorescent signals. These advances should substantially expand the capability to query parent-of-origin-specific chromosome positioning and gene expression on a cell-by-cell basis.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Haplótipos
/
Cromossomos
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Hibridização in Situ Fluorescente
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Coloração Cromossômica
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Análise de Sequência com Séries de Oligonucleotídeos
Limite:
Animals
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article