Today Anthony and I started preparation for DDW and D2O experiments with yeast and e. coli. Anthony has a few posts about our past growth experiments. Previously we were successful growing both organisms however, we didn’t get into the DDW or D2O phase of the experiments.
The yeast we are using is S. cerevisiae, g160/2d. We are growing three samples. One sample is being grown in YPD broth that is a mixture of 50mL SIGMA Db/processed water and 2.5g of USB YPD Broth (ultrapure). The other two samples are on 100mm YPD Agar plates from Teknova. One plate is in the incubator with the broth mixture at 24 degrees C. The other plate we are leaving out at room temperature.
We are growing two samples of DH5α™ e. coli. The first sample on a 100mm LB Agar plate (also from Teknova). The second is in an LB broth mixture of 1g LB broth mixed with 50mL of Dbl processed water, both from SIGMA. Both samples are in the incubator at 37 degrees C.
We made LB and YPD broth by carefully measuring the indicated amount of broth powder using the lab’s Ohause Adventurer SL balance and small weight boats. We then poured the powder from the weight boat into 50mL beakers and added 50mL of water. We then used magnetic stirrers and a hotplate/stirrer to mix both broths. The LB broth was autoclaved.
Tomorrow we should have some samples ready to go. We are planning on first seeing if we can grow deuterium resistant strains of both the yeast and e.coli. After that we will see how the resistant strains fair in DDW.