Characterization of synthetic DNA bar codes in Saccharomyces cerevisiae gene-deletion strains.
Proc Natl Acad Sci U S A
; 101(30): 11046-51, 2004 Jul 27.
Article
em En
| MEDLINE
| ID: mdl-15258289
ABSTRACT
Incorporation of strain-specific synthetic DNA tags into yeast Saccharomyces cerevisiae gene-deletion strains has enabled identification of gene functions by massively parallel growth rate analysis. However, it is important to confirm the sequences of these tags, because mutations introduced during construction could lead to significant errors in hybridization performance. To validate this experimental system, we sequenced 11,812 synthetic 20-mer molecular bar codes and adjacent sequences (>1.8 megabases synthetic DNA) by pyrosequencing and Sanger methods. At least 31% of the genome-integrated 20-mer tags contain differences from those originally synthesized. However, these mutations result in anomalous hybridization in only a small subset of strains, and the sequence information enables redesign of hybridization probes for arrays. The robust performance of the yeast gene-deletion dual oligonucleotide bar-code design in array hybridization validates the use of molecular bar codes in living cells for tracking their growth phenotype.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
/
DNA Fúngico
Idioma:
En
Revista:
Proc Natl Acad Sci U S A
Ano de publicação:
2004
Tipo de documento:
Article
País de afiliação:
Estados Unidos