A Saccharomyces cerevisiae cell-based quantitative β-galactosidase assay compatible with robotic handling and high-throughput screening

Rogerio Alves de Almeida, Danielle Burgess, Reut Shema, Nuzhat Motlekar, Andrew D. Napper, Scott L. Diamond, Graham D. Pavitt

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Reporter-gene assays that employ the Escherichia coli lacZ gene are ubiquitously employed in biological research. However, we were not able to readily identify a quantitative method that worked reliably with yeast (Saccharomyces cerevisiae) cells and that was compatible with high-throughput screening and robotic liquid handling tools. We have therefore adapted a commercially available assay employing a 6-O-β-galactopyranosyl-luciferin substrate to provide the required sensitivity with minimal sample handling times. Our assay uses only one-tenth of the reagents suggested by the reagent manufacturer (Promega) for equivalent assays with mammalian cell cultures and produces rapid, sensitive and reproducible analysis with as little as 1 μl yeast cell culture and with
    Original languageEnglish
    Pages (from-to)71-76
    Number of pages5
    JournalYeast
    Volume25
    Issue number1
    DOIs
    Publication statusPublished - Jan 2008

    Keywords

    • β-galactosidase
    • High-throughput screening
    • LacZ reporter assay luciferase

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