Evan’s Capstone – eNotebook 2

Tuesday – 1/26/2016

Coho Salmon and Cutthroat Trout Liver Analyses

Continuing work on RNA isolations today, managed to double up on the samples run per day and actually ended up easier to do than Thursday’s due to running homogenization during incubation steps.

Samples were pulled from Swamp Creek and May Creek again, Set 1 being samples 2May, 6May, 9Swamp, 14Swamp at 11:36 AM; Set 2 consisted of 1May, 7May, 6Swamp, and 18Swamp and were pulled at 12:54 PM.

Samples were ran through RNA Isolation as indicated in the Common Lab Protocols like last Thursday’s samples (found at https://github.com/sr320/LabDocs/wiki/Common-Lab-Protocols).

Some more minor deviations due to timing between sets, less than before given overlap went pretty well overall during incubation times. The 1st set of samples were kept on ice from 2:30 PM to 3:30 PM until time was available to do the Nanodrop. The 2nd set of samples had to sit at Step 13 for 2 extra minutes, and on Step 10 for 3 minutes, then before Step 20 they had to remain in the isopropanol for 10 minutes while waiting for Nanodrops for the 1st set.

Nanodrop samples came back better than before (likely due to being more comfortable with the machine) and were conducted the same way as Thursday’s samples.

Set 1 ~
2BMA – 260/280: 1.99; 260/230: 2.14; Concentration: 1466.4
6BMA – 1.84, 1.87, 422.4
9BSW – 1.98, 2.12, 1505.1
14BSW – 1.98, 1.86, 1387.5

Set 2 ~
1BMA – 1.96, 1.82, 889.9
7BMA – 2.00, 2.10, 1841.1
6BSW – 1.98, 2.14, 1410.4
18BSW – 1.99, 1.82, 1699.6

Samples in Set 1 were placed in the -80 Celsius freezer at 3:50 PM while the Set 2 samples were in the freezer by 4:50 PM.

Seems to be even better than last week, plan for Thursday is to learn the process of DNasing the RNA samples and likely more RNA isolations!