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

• Type: Article in International Scientific Journal
• Date: 1999

Authors

Mergulhao, FJM (Author) Other; Kelly, AG (Author) Other; Monteiro, GA (Author) Other; Taipa, MA (Author) Other; Cabral, JMS (Author) Other

Journal

Title: Molecular Biotechnology

Vol. 12

Pages: 285-287

ISSN: 1073-6085

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 6 Citations

ISI Web of Science

Scopus - 9 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

CORDIS: Technological sciences > Technology > Biotecnology

Other information

Authenticus ID: 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 3