Cait Barrett Entry #3 Liquid Nitrogen

Cait Barrett Entry #3, Liquid Nitrogen

This past week Ben and I had to make new RNA for his course he is teaching at Woods Hole. The course is on injections, and the RNA is used to rescue, or bring back to normal state, embryos which have been injected with morpholinos that causes either a dorsal or ventral phenotype. To make these RNAs we had to use bacteria with our plasmid in it. This caused us to be culturing bacteria on plates, and to do this we rolled the bacteria out onto plates with ampicillin. The ampicillin in the plates kills bacteria that are not resistant, so bacteria with our plasmid that have resistance would survive while random bacteria would not. After we grew colonies on this plate we placed those bacterial colonies, one in each tube of the different plasmid forms, into tubes filled with a substance called Terrific Broth. As the name implies, it was terrific at growing our bacteria and we had plenty to purify using a protocol of Isopropanol washes and enzymes that cut our plasmid in order for only the specific gene we wanted to be transcribed.

After all this was done we were able to set some of the bacteria aside to be used in the future. This process involved snap freezing, which is when you freeze something extremely rapidly. We mixed our bacteria with glycerol in order to preserve it, and then placed those tubes into a bucket of Liquid Nitrogen. A fun fact about liquid nitrogen is that as it leaves the container it makes the most outrageous and sci-fi movie-esque sounds. We then placed these tubes into Ben’s box that goes in the -80 degrees Celsius freezer.

Also during this week Matt came into Philly and we were able to catch a Phillies game. Although I am not a fan it was fun to do something in Philly outside of just work. Plus the Phillies lost, so I could not complain.