Troubleshooting in gene splicing by overlap extension: A step-wise method

• Tipo: Artigo em Revista Científica Internacional
• Data: 1999

Autores

Mergulhao, FJM (Autor) Outra; Kelly, AG (Autor) Outra; Monteiro, GA (Autor) Outra; Taipa, MA (Autor) Outra; Cabral, JMS (Autor) Outra

Revista

Título: Molecular Biotechnology

Vol. 12

Páginas: 285-287

ISSN: 1073-6085

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 6 Citações

ISI Web of Science

Scopus - 9 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Ciências biológicas

CORDIS: Ciências Tecnológicas > Tecnologia > Biotecnologia

Outras Informações

ID Authenticus: P-007-8W3

DOI: 10.1385/mb:12:3:285

Abstract (EN): A step-wise method for cloning intron-containing genes from genomic DNA is described. The two exons of the human proinsulin gene were separately amplified in two steps using, in the first step, completely homologous primers. This reduces unwanted interactions between mismatched primers and a complex DNA template such as genomic DNA. The fragments were amplified in a second step polymerase chain reaction (PCR) using mismatched primers that incorporated additional bases complementary to the other exon, and these products were spliced together in a third step PCR.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 3