Second batch of 24 Crab hemolymph samples - RNA extracted and qubit results

Today I did another round of Day 9 infected (12) and uninfected (12) tanner crab hemolymph RNA extractions. I also ran 2ul of the eluted 15ul RNA on Qubit. Results and details in post.

RNA extraction

I followed yesterday’s (post) extraction process as exactly as I personally could. Nothing went wrong as far as I’m aware…

img

Some notes:

The following tubes had small amounts of initial pelleted hemolymph:
71, 67, 177, and 44

img

The following tubes had white crystal-y stuff in them after sample preparation step:
20, 54, 24, 169, 29

img

After Step 2 (running ethanol through Zymo yellow column), tubes 44 and 29 didn’t fully run through, so I ran them through the centrifuge one more time at 10,000 g for 30 sec.

I saved the DNA again, and put with yesterday’s saved DNA in FTR 209, -20C on the labeled tube rack.

Results

Raw qubit file
Google spreadsheet with tube-numbers

Just like with yesterday’s batch of extracted RNA, I ran 2ul of each sample on Qubit with RNA HS kit.

23/24 tubes had detectable RNA!!!! WOOOOOO!!!

qubit_tube_conc_ng.ml original_sample_conc_ng.ul sample_vol_ul dilution_factor tube_number extraction_method ul_sample-used elution_vol_ul total-yield_ng
258 25.8 2 100 29 Zymo_microprep 35 15 335.4
Out of range Out of range 2 100 44 Zymo_microprep 35 15 #VALUE!
130 13 2 100 1 Zymo_microprep 35 15 169
228 22.8 2 100 177 Zymo_microprep 35 15 296.4
115 11.5 2 100 95 Zymo_microprep 35 15 149.5
166 16.6 2 100 124 Zymo_microprep 35 15 215.8
139 13.9 2 100 67 Zymo_microprep 35 15 180.7
118 11.8 2 100 7 Zymo_microprep 35 15 153.4
464 46.4 2 100 169 Zymo_microprep 35 15 603.2
212 21.2 2 100 24 Zymo_microprep 35 15 275.6
117 11.7 2 100 11 Zymo_microprep 35 15 152.1
134 13.4 2 100 5 Zymo_microprep 35 15 174.2
147 14.7 2 100 166 Zymo_microprep 35 15 191.1
174 17.4 2 100 17 Zymo_microprep 35 15 226.2
231 23.1 2 100 117 Zymo_microprep 35 15 300.3
145 14.5 2 100 102 Zymo_microprep 35 15 188.5
186 18.6 2 100 151 Zymo_microprep 35 15 241.8
231 23.1 2 100 170 Zymo_microprep 35 15 300.3
840 84 2 100 172 Zymo_microprep 35 15 1092
186 18.6 2 100 158 Zymo_microprep 35 15 241.8
252 25.2 2 100 54 Zymo_microprep 35 15 327.6
277 27.7 2 100 20 Zymo_microprep 35 15 360.1
100 10 2 100 63 Zymo_microprep 35 15 130
182 18.2 2 100 71 Zymo_microprep 35 15 236.6

Samples are in -80 along with the extracted samples from yesterday (Rack 7, column 3, row 4)

The remaining pelleted hemolymph are in the same box as yesterday’s (Rack 8, column 3, row 5)

Next steps:

On Monday, I’d like to run these samples on the bioanalyzer and/or Nanodrop to make sure that things look good.

Written on October 25, 2019