DNA for Competition 1 was extracted from each sample using a protocol modified from Hoffman (2002 ) that included adding approximately 200 μl of 0.5 mm diameter glass beads (BioSpec Products, Bartlesville, OK) to each sample and lysing cells in a bead beater (BioSpec Products) on high for 5 min at room temperature. For samples grown in Grape, DNA was also column purified to remove an unknown inhibitor of PCR amplification. DNA for Competition 2 was extracted using the ZR‐96 Fungal/Bacterial DNA Kit (Zymo Research, Organge, CA).
0.5 mm diameter glass beads
Manufactured by Biospec
The 0.5 mm diameter glass beads are a precise, uniform, and high-quality laboratory equipment item. They are made of durable glass material and measure 0.5 mm in diameter. These glass beads can be used for various laboratory applications that require small, spherical particles.
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2 protocols using 0.5 mm diameter glass beads
1
DNA Extraction for Competition Samples
2
Yeast Cell Lysis and Western Blot
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Yeast cells (approximately 12 OD600) were pelleted, washed with sterile water, and lysed in 100 μl SDS-sample buffer with 0.5 mm diameter glass beads (BioSpec Products, Bartlesville, OK) by two rounds of boiling for 2 min and vortexing for 30 sec. Protein extracts were centrifuged at 13,000 rpm for 15 min, and the supernatants analysed by Western blot. Western blot signals were developed using ECL (GE Healthcare, Little Chalfont UK).
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