Inositol Labelling of Yeast Cells: Difference between revisions
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# Aspirate the supernatant | # Aspirate the supernatant | ||
# Wash the cells one time in 1 ml inositol-free media (resuspend cells, spin at 14,000RPM for 1 min, and aspirate supernatant) | # Wash the cells one time in 1 ml inositol-free media (resuspend cells, spin at 14,000RPM for 1 min, and aspirate supernatant) | ||
# Resuspend the cells in 875 | # Resuspend the cells in 875 uL inositol-free media (2 OD/ml) | ||
# At 10:00 PM inoculate each of the three 50 ml Falcon tubes with the appropriate amount of each cell type based on the following considerations | # At 10:00 PM inoculate each of the three 50 ml Falcon tubes with the appropriate amount of each cell type based on the following considerations | ||
## The amount for inoculation will vary based on the growth rate of each cell strain (Wild type cells need about 60 uL) | ## The amount for inoculation will vary based on the growth rate of each cell strain (Wild type cells need about 60 uL) | ||