Analysis of cosmids using linearization by phage lambda terminase.
Gene
; 40(2-3): 259-66, 1985.
Article
em En
| MEDLINE
| ID: mdl-3007292
ABSTRACT
A group of cosmid clones was isolated from the region of the mouse t complex and analysed by a rapid restriction mapping protocol based on linearization of circular cosmid DNA in vitro. A plasmid capable of producing high levels of phage lambda terminase was constructed and procedures for in vitro cleavage of cosmid DNAs were optimised. After linearization, the cosmids were partially digested with restriction enzymes, and either cos end was labelled by hybridization with radioactive oligos complementary to the cohesive end sequence, a step which we have described previously for clones in phage lambda (Rackwitz et al., 1984). High-resolution restriction maps derived by this method were used to identify and align the cosmids, to localise the position of repetitive sequences, and to interpret the results of electron microscopy heteroduplex experiments.
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Base de dados:
MEDLINE
Assunto principal:
Cosmídeos
/
Bacteriófago lambda
/
Desoxirribonucleases
/
Endodesoxirribonucleases
/
Endonucleases
/
Escherichia coli
Limite:
Animals
Idioma:
En
Revista:
Gene
Ano de publicação:
1985
Tipo de documento:
Article