Evan’s Capstone – eNotebook 4

Thursday – 2/4/2016
            Back in the lab after calling in sick on Tuesday, have to make up time tomorrow! Just continuing the RNA isolations, should be done with the last 7 tomorrow (leaving me with a total of 35 samples between 6 watersheds).

Samples ran today was the last set to finish off Harris and Issaquah from Thursday (2 and 19 from Issaquah in addition to 11 and 16 from Harris) and those were pulled at 12:45 PM. The other set of samples was the beginning of Coulter Creek and Rock Creek (9BCO, 14BCO, 6BRC, and 7BRC) and were pulled at 1:30 PM.

Surprisingly, there was actually no deviations today at all from the common procedure as all the livers were of a large enough size in addition to having the lab to myself. The optimal strategy seems to be to start homogenization of the 2nd set while the 1st set is incubating for the first centrifuge step. It ends up leaving you swapping out samples constantly with there always being one set of samples in the centrifuge at any point in time!

Nanodrop Results for the Day:

2BIS – 260/280: 1.94, 260/230: 2.19, Concentration: 769.1 ng/ul
19BIS – 1.80, 2.12, 435.5
11BHA – 1.98, 2.19, 1265.6
16 BHA – 2.00, 2.11, 1473.3

6BRC – 1.97, 2.18, 1276.7
7BRC – 1.96, 2.21, 1860.0
9BCO – 1.92, 2.21, 825.7
14BCO – 1.77, 2.03, 446.5

I also seems like I finally figured out the Nanodrop machine, getting results on the first attempt for all but the Coulter Creek samples which was giving me errors. I later learned (Thanks Sam!) that the machine seems to respond well to 2 ul samples instead of 1 ul samples, as it seems to prevent column breakage which I think is what has been giving me such a hard time for the last 3 tries at the machine. Gives me a little more hope for the samples from last Thursday that maybe I simply needed to increase the water drop size for the machine. Would have liked better results from 19BIS and 14BCO, but they are pretty close and the concentrations are lower than the other samples so it is likely fine.

All samples were placed back in the -80 freezer at 5:10 PM and I’m going to try to work in some time tomorrow to get the last 7 samples done to finish off the week!