Growth Curve Experiment

Chemical transformation of yeast requires the organism to be in logarithmic growth phase, so, before we clone our casein genes into yeast, we need to make sure that we know how to grow our yeast to log Phase.

Here's a brief protocol for monitoring yeast growth.

• GROWTH CURVE

(1) Figure out growth curve of yeast strains (we have Carolina strain now, we want ATCC strain, and Keoni Strain)

    (a) please be conservative with YPD media here - we can do these experiments in 100mL           
         tubes - in triplicate.  Record the OD (650nm) at 0, 5, 10, 24, 48, and 72 hours. 

(2) Repeat growth curve with pD1214:FAKS-hKappa cloned into yeast strain.

    (a) no, it's not the real human kappa casein, but it'll get us closer to understanding how our strains will perform after real cloning happens. 
         This is a great set of experiments for folks who've not done cloning before. Please learn about Bacterial transformation
         and plating for single colonies, Plasmid Prep, and Yeast transformation.
    (b) grow the yeast clones in triplicate (100mL) to create growth curve.

Select a single colony and grow it overnight in 5mL YPD at 30C, rotating at 250rpm. Read the OD the next morning (>16hrs later). Record the OD at 16hours, then start the culture with a KNOWN amount of 16-hr innoculum.

     (a) Dilute the 16hour overnight in YPD, or 0.1mL into 99.9mL (1:1000) YPD.

     (b) Record the OD (650nm) at 0, 5, 10, 24, 48, and 72 hours.