A Test Bed for Budding Technologies

DELETION BY DESIGN:Courtesy of Guci GiaeverThe deletion cassette module used to delete each yeast gene contains two 74-basepair tags upstream and downstream (UPTAG and DNTAG) of the KanMX gene, which confers resistance to the drug geneticin. UPTAG and DNTAG contain 18 basepairs of genomic sequence to flank the yeast's open reading frame, and U1 and U2, or D1 and D2 PCR primers for amplifying a unique 20-basepair TAG region-the so-called molecular barcode. A second round of PCR adds 45 base-pairs

Written byAileen Constans

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Courtesy of Guci Giaever

The deletion cassette module used to delete each yeast gene contains two 74-basepair tags upstream and downstream (UPTAG and DNTAG) of the KanMX gene, which confers resistance to the drug geneticin. UPTAG and DNTAG contain 18 basepairs of genomic sequence to flank the yeast's open reading frame, and U1 and U2, or D1 and D2 PCR primers for amplifying a unique 20-basepair TAG region-the so-called molecular barcode. A second round of PCR adds 45 base-pairs of ORF-specific sequence to either end of the cassette (UP_45 and DOWN_45), enhancing the specificity during homologous recombination.

Beyond its roles in beer and bread production, yeast has come into its own as a model organism for large-scale studies. Used famously to piece together the rudimentary components of eukaryotic cell-cycle processes, this simple organism with its quick life cycle and facile genetics continues to be the prototype for creating functional genomics ...

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