Today in lab we altered YPD media to pH 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and confirmed with a digital pH meter. We altered 10 mL of YPD solution which began at a pH of roughly 6.3 to each desired pH. We did this by adding 1M NaOH and 1M HCl drop by drop until we reached desired pH.
The digital pH reader confirmed pHs as: 1.99, 2.98, 3.96, 5.00, 6.03, 7.05, 7.99, 8.99
After YPD media was changed each pH was split into 3 for labeling and inoculation with wild strain yeast, TOS3 deficient yeast, and SAK1 deficient yeast.First we will to dilute the yeast sample at 1/32 by adding 4uL to 126uL of YPD media. We are reducing to 3 replicates for financial reasons.
130 mL of Inoculated YPD media was added to each well on a 96 well plate as explained in our procedure ((see previous lab post for chart of 96 well plate)).
note: A drop of yeast spilled between well 7A and 8A. and possibly others.