S-STEM Scholar Kristi Morrison's Blog

The extraction of G6PD was performed by centrifuging 100 ml of seven different bacteria; Deinococcus indicus, Deinococcus radiodurans, Deinococcus caeni, Deinococcus pimensis, Deinococcus hopiensis, Deinococcus grandis, Deinococcus metalli. We then resuspend the cell pellets in 500 ul extraction buffer. The extraction buffers keep G6PDH enzyme in an active state. Next, we put the tubes in Fisher Scientific BeadMill and ran the device for four rounds at 5 m/sec for 90 seconds, with a 1 minute ice bath in between each round. We ended with transferring the supernatant to a new Eppendorf tube with 200ul pipette.

This week I made 8 250ml TGY media. 7 different bacteria were inoculated at 28 degrees celsius. They were then placed in the incubator shaker for 24 hours.  Cell culture growth was measured at 600nm absorbance. I used the spectrometer to make sure the bacteria had a 0.1 absorbance. This step was taken to ensure that the bacteria was in their exponential growth phase.