S-STEM Scholar Rebecca Matadamas's Blog

During this lab, we were subculturing. We put aquaticus German in TGY, Caenie in R2A, Deserti in TSB/10, Hopi in TGY, Grandis in PYEA, Pima in R2A, D. Rad in TGY, Sonoraensis in R2A, Indicus in TSYE, and Geothermalis in NA. It went well and now we wait for them to grow.

On 4/14, we were prepping plasmid. Trevon and Laura were prepping the PRad and Shiva and I prepped the PRadZ1. At first, the centrifuge was being questionable, so after rescuing our samples from that centrifuge, we switched them to a more reliable one. In the end, our sample was pretty dirty, at 8.7 and 8.8 concentration, and 2.17 and 2.69 260/280 ratio.

On 4/2 I performed my first gram stain. It was nice to actually do something that is talked about so casually in the lab. I gram stained 3 different samples, two aquaticus and one fluorescens. This picture is of the fluorescence sample, showing it to be gram negative. On 4/6, we did an inoculation of transformed e. coli to prep for our 4/7 plasmid prep/isolation. On 4/7 we did out plasmid prep, and one of our samples from the day before did not grow, so we were pulling from only one sample. By the end, we saw that our samples were pretty dirty, having a 260/280 ration of 2.21 and 2.08, but it was expected, being our first prep.

During this lab, we really went into depth on using micro-pipettes. From my last post, I had never touched a micro-pipette, let alone know how to use one. There are 4 micro-pipettes that we use, P-20, P-100, P-200, P-1000. Their names correspond with the maximum volume they can hold. When using a micro-pipette, you do not want to push all the way down when collecting a sample, or else it will pick up its maximum volume. Push down to pick up the sample, then slowly lift your finger from the button to retrieve. The P-1000 is equivalent to 1 mL and it takes the big tips while the other three pipettes take the small tips. I learned a lot about these pipettes, but I think one of the most important pieces I learned was to always distribute the larger volume of your sample first, then mix your smaller volume in.