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) Streak Carolina strain on a YPD + Ura plate for isolated colonies.
    (b) Grow overnight at 30C
    (c) Pick a single colony from the plate (in triplicate) and grow in 5mL YPD + Ura
    (d) The next day, Record the OD (650nm) of the >16hour culture, and dilute the Growth Curve Culture by adding 100uL of 16hour culture to 99.9mL of YPD + Ura.
    (e) Grow at 30C, 250rpm.
    (e) Record the OD (650nm) at 0, 5, 10, 24, 48, and 72 hours. Use YPD + Ura as the blank for each reading.

(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) Transform positive plasmid into E. coli (see Molecular Biology page)
    (c) Instead of plating transformation, add the transformation reaction to 5mL of LB + Carb(50).
    (d) Grow this plasmid overnight at 37C, rotating; and plasmid prep the DNA the next day.
    (e) Transform yeast using the Yeast Transformation protocol.
    (f) Plate transformation and incubate overnight at 30C.
    (g) Select single colonies and repeat from (1) - (c) (above).

(3) RECORD YOUR DATA, MAKE A FIGURE, and POST IT HERE.