An efficient and site-specific gene trimming method.
Biotechniques
; 7(5): 506-10, 1989 May.
Article
em En
| MEDLINE
| ID: mdl-2699239
ABSTRACT
Trimming a DNA strand into a precisely determined fragment can be carried out efficiently by an improved method involving a site-specific trim-primer and a single-stranded DNA template which is generated from a multifunctional vector, pTZ18R, and linearized by using an Eco RI-pTZ18R splinter. A complementary DNA strand is synthesized by DNA polymerase I (Klenow), and the 3'-end of the template upstream from the annealed primer is trimmed by subsequent T4 DNA polymerase reaction. An ATG translation initiator codon or a termination codon can be incorporated into the trim-primer, providing versatility to this single-stranded DNA-initiated gene trimming method that can be applied to subcloning and expression of any DNA fragment with known terminal sequences.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Técnicas Genéticas
Limite:
Humans
Idioma:
En
Ano de publicação:
1989
Tipo de documento:
Article