Simultaneous shotgun sequencing of multiple cDNA clones.
DNA Seq
; 7(2): 63-70, 1997.
Article
em En
| MEDLINE
| ID: mdl-9063643
ABSTRACT
This study describes an efficient method for the rapid sequencing of DNA fragments in the size range 1-5 kb. Individual fragments, here cDNA inserts, are purified by restriction enzyme digestion and gel purification, pooled and concatenated by ligation. The concatamers are sheared and cloned randomly into M13, followed by random sequencing. The sequences of the individual cDNA inserts are obtained at the assembly stage using restriction enzyme sites as "tags' for the ends of each fragment. In this study the sequencing of two libraries containing 7 and 16 cDNA inserts with an average length of 2.5 kb is described. The results show that of the shotgun sequencing of large fragments, and that the method compares favorably to alternative strategies, such as primer walking.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Clonagem Molecular
/
Análise de Sequência de DNA
/
DNA Complementar
Limite:
Animals
Idioma:
En
Revista:
DNA Seq
Assunto da revista:
BIOLOGIA MOLECULAR
/
BIOQUIMICA
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Estados Unidos