微染色体－微管结合实验（Minichromosome

相关专题   MMB MINICHROMOSOME-MICROTUBULE BINDING ASSAY Cell Determine the OD600 and correlate the density from the chart. Set up four 100mL YPD cultures at the following densities: 0.7x105, 1x105, and 3.0x105 cells/mL GROWTH OF CELLS Grow 100mL of cells to OD600=0.7-0.8 at 23oC. Add 5-10ul of BME, 15 minutes before spin. Harvest cells in two 50mL conical tubes, spin 1.5-2.0K for 8-10min. SPHEROPLASTING. Resuspend cells in 4mL YWB. *NOTE: If cells have been arrested with nocodazole or alpha factor, include the inhibitor in the YWB. Let sit at room temperature for 2-3 min. Add oxalyticase