In situ genotyping individual DNA molecules by target-primed rolling-circle amplification of padlock probes.
Nat Methods
; 1(3): 227-32, 2004 Dec.
Article
em En
| MEDLINE
| ID: mdl-15782198
ABSTRACT
Methods are needed to study single molecules to reveal variability, interactions and mechanisms that may go undetected at the level of populations of molecules. We describe here an integrated series of reaction steps that allow individual nucleic acid molecules to be detected with excellent specificity. Oligonucleotide probes are circularized after hybridization to target sequences that have been prepared so that localized amplification reactions can be initiated from the target molecules. The process results in strong, discrete detection signals anchored to the target molecules. We use the method to observe the distribution, within and among human cells, of individual normal and mutant mitochondrial genomes that differ at a single nucleotide position.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Mitocondrial
/
Sondas de DNA
/
Análise de Sequência de DNA
/
Hibridização in Situ Fluorescente
/
Marcação de Genes
/
Técnicas de Amplificação de Ácido Nucleico
/
Doenças Mitocondriais
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Nat Methods
Assunto da revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Ano de publicação:
2004
Tipo de documento:
Article
País de afiliação:
Suécia