Generation of clonal DNA templates for in vitro transcription without plasmid purification.
Biotechniques
; 8(3): 252-7, 1990 Mar.
Article
em En
| MEDLINE
| ID: mdl-2184850
ABSTRACT
WE present a rapid procedure based on the polymerase chain reaction for generation of double-stranded DNA templates suitable for in vitro transcription by T3 or T7 RNA polymerase. DNA fragments cloned into a phage promoter vector are amplified together with a flanking promoter to provide functional templates. Extension of oligonucleotide primer molecules harboring an RNA polymerase promoter sequence at their 5'-end allows positioning of the transcription start site within the insert. The procedure generates large amounts of linear transcription template without need to isolate and purify plasmid DNA from bacterial cells.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Transcrição Gênica
/
DNA Viral
/
Técnicas Genéticas
Tipo de estudo:
Etiology_studies
/
Observational_studies
/
Risk_factors_studies
Idioma:
En
Revista:
Biotechniques
Ano de publicação:
1990
Tipo de documento:
Article