Week 3: Growing things

Returning Monday from a week-long absence due to cross country camp and the ASE Midsummer conference, I found myself forgetting where a few things are around the lab. Roberto took every opportunity to poke fun at me for it, say that my brain had "gone to mush" while I was gone. Jokingly, of course.

My week has consisted almost entirely of making media and then growing things on those media. On Monday, I made some X media on which to grow Halomonas LOB-5. On Wednesday, we pulled it out from the -80°C freezer, where I stupidly grabbed a shelf for too long without the big blue freezer gloves, and plated it onto X media and some K media that I had made a previous week. They were incubated at 10°C and 30°C. Currently, the ones growing on K media are growing a bit faster than those on X at their respective temperatures and the ones growing at 30°C are growing much faster than those at 10°C. After 41 hours of incubation, there are single cultures on K media that has been at 30°C and I probably go subculture right after I finish writing this.

On Tuesday, I had the pleasure of making more media, this time with Columbia River Plume water as a base. This would be used to culture bacteria from the PC2 experiment that Roberto observed to be oxidizing Mn after 2 months at 10°C while I was gone. The water had to be filtered to sterilize it so that nothing would precipitate out, as it might have had it been autoclaved. After filtering the water, we removed the filters and subjected them to an LBB assay, out of curiosity to see whether there were MN-oxides in the water. Fortunately, the filters tested positive, confirming Roberto's observations of the bacteria. Concentrated agar was made and autoclaved with a few other things mixed in and the water was added directly before pouring plates so that it wouldn't cool the agar too much. I made three different media with this: K, Lept, and a minimal medium that Roberto calls CRMM.  

Also, on Tuesday, I discovered at the lab meeting that Amy and I will have to present our talks to the lab on August 3rd, more than 2 weeks before they have to be ready for the ASE Symposium, so that should be pretty fun...

Yesterday, around 3pm, I subcultured one of those bacteria mentioned above onto Lept media and, by this morning, there were already single cultures visible on the plate that was incubated at room temperature. I still have a lot more subculturing to do today as we try to purify bacteria from the PC2 experiment.