Isoform discovery by targeted cloning, 'deep-well' pooling and parallel sequencing.
Nat Methods
; 5(7): 597-600, 2008 Jul.
Article
em En
| MEDLINE
| ID: mdl-18552854
ABSTRACT
Describing the 'ORFeome' of an organism, including all major isoforms, is essential for a system-level understanding of any species; however, conventional cloning and sequencing approaches are prohibitively costly and labor-intensive. We describe a potentially genome-wide methodology for efficiently capturing new coding isoforms using reverse transcriptase (RT)-PCR recombinational cloning, 'deep-well' pooling and a next-generation sequencing platform. This ORFeome discovery pipeline will be applicable to any eukaryotic species with a sequenced genome.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Clonagem Molecular
/
Análise de Sequência
/
Isoformas de Proteínas
Limite:
Animals
/
Female
/
Humans
/
Male
/
Pregnancy
Idioma:
En
Revista:
Nat Methods
Assunto da revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Estados Unidos