Notes 20100722 Rohan's Talk

From SnOwy - Ed's Wiki Notebook

biotechnology paper

• rapid profiling, microbial genome, mixtures barcoded oligonucleotides
• high throughput genome manipulation
  • do all of the tedious things automatically
  • do the actual manipulation very quickly
• how did we figure out the minimal yeast genome?
  • replaced every ORF with a KAN-R (Kanomycin Resistance) marker
    • now in general called a molecular barcode
• deletion strains
  • specific markers knocked out
  • auxotrophic markers (ADE, TRP) used
    • knockouts are less viable in a nutrient restricted medium
• implies we are less able to harvest such individuals
• recombineering
  • insertion of synDNA cassette rather than site mutation
  • multiplex automated genome engineering (MAGE)

Trackable Multiplex Recombineering (TRMR)

• combines the following technologies
  • barcoding (tagging)
  • synthetic DNA constructs
  • recombineering
  • microarray analysis
• creates high-throughput system -- alters genomes quickly; low cost, deduces underlying trait without prior knowledge of biology
• allows enrichment of improved cells
  • enrichment means filtering out less successful strains (mutants)