Sequential amplification of flanking sequences by Y-shaped adaptor dependent extension using multiple templates.
Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao
; 33(1): 85-90, 2007 Feb.
Article
em En
| MEDLINE
| ID: mdl-17287574
ABSTRACT
Y-shaped adaptor dependent extension (YADE) method is a useful tool to amplify the flanking sequence of a known DNA sequence, but its efficiency is frequently limited by the restriction sites around the known sequence. In this paper, we demonstrated that using multiple templates derived from several restrictions and ligations could dramatically increase the efficiency of YADE method and render it suitable for sequential amplification of flanking sequences. With templates originating from 7 digestions, a 2,228-bp 5'-upstream sequence of a cotton small GTPase gene was obtained by two rounds of sequential YADE amplifications. The results demonstrated that the YADE method with multiple templates may be a useful tool for sequential PCR walking in complex genomes.
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Base de dados:
MEDLINE
Assunto principal:
Moldes Genéticos
/
Reação em Cadeia da Polimerase
/
DNA de Plantas
Idioma:
En
Ano de publicação:
2007
Tipo de documento:
Article