Application: DNA/RNA and Other Extractions
The wealth of information generated from years of biochemical, genetic, and molecular analyses has made yeasts both model biological systems and tools for bio pharmaceutical scientists. Consequently, yeast are a popular host for gene expression studies and for the production of recombinant proteins. Though many yeast species are in use, including Pichia, Hansenula, and Debaryomyces, the most popular yeast continues to be Saccharomyces. Yeast mRNA and intracellular proteins are often times difficult to extract intact from cells by traditional enzymatic methods. Lysing enzymes are often crude preparations containing RNase and proteases that will not only attack the cell wall, but also the molecules of interest. Furthermore, protoplasts generated from enzymatic digestion usually require lysis with detergents that will also denature many proteins to inactivity. Therefore, mechanical cracking/fracturing of the yeast cell is often required to liberate the molecules.
Mechanical disruption of yeasts has traditionally been accomplished by using either a press or bead mill (i.e., bead beater). In both approaches, samples are processed individually. For experiments where large numbers of yeast clones must be examined in a high-throughput screening environment, individual processing is a major bottleneck and impractical. Consequently, a method is needed that combines mechanical disruption of cells in a high-throughput format. The Geno/Grinder, a bead mill originally designed to smash seeds in deep well plates, can be used to disrupt yeast in a microwell plate format.
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