Optimized multiplex PCR: efficiently closing a whole-genome shotgun sequencing project.
Genomics
; 62(3): 500-7, 1999 Dec 15.
Article
em En
| MEDLINE
| ID: mdl-10644449
ABSTRACT
A new method has been developed for rapidly closing a large number of gaps in a whole-genome shotgun sequencing project. The method employs multiplex PCR and a novel pooling strategy to minimize the number of laboratory procedures required to sequence the unknown DNA that falls in between contiguous sequences. Multiplex sequencing, a novel procedure in which multiple PCR primers are used in a single sequencing reaction, is used to interpret the multiplex PCR results. Two protocols are presented, one that minimizes pipetting and another that minimizes the number of reactions. The pipette optimized multiplex PCR method has been employed in the final phases of closing the Streptococcus pneumoniae genome sequence, with excellent results.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase
/
Genoma Bacteriano
/
Análise de Sequência de DNA
/
Técnicas de Química Combinatória
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
Idioma:
En
Revista:
Genomics
Assunto da revista:
GENETICA
Ano de publicação:
1999
Tipo de documento:
Article
País de afiliação:
Estados Unidos