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Characterization of synthetic DNA bar codes in Saccharomyces cerevisiae gene-deletion strains.

Authors
  • Rg, Eason
  • Nader Pourmand
  • W, Tongprasit
  • Zs, Herman
  • K, Anthony
  • O, Jejelowo
  • Rw, Davis
  • V, Stolc
Type
Published Article
Journal
Proceedings of the National Academy of Sciences
Publisher
Proceedings of the National Academy of Sciences
Volume
101
Issue
30
Source
UCSC Bioengineering biomedical-ucsc
License
Unknown

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.

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