Protein Content of Madracis decactis and Diploria labyrinthiformis tissue for Physiological Analyses

This post entails the date each sample was processed to determine protein concentration using the Peirce Rapid Gold BCA assay kit. See HERE for the protocol

20241206

• Samples were all pratice samples used for protocol practice. |Samples| Box| |——-|—-| |DLextra| 5| |MD-1-31| 5 | |DL-4-33|5| |MD-4-35|5| |MD-1-36|5|

(9 standards + 5 samples) x (2 replicates) x (200 μL of WR) = 5,600μL = 5.6mL WR (Prepare 7) 50 Reagent A:1 Reagent B 7mL:0.14mL

20241207

|Samples| Box| |——-|—-| |DLextra| 5| |DL-4-33|5| |MD-4-35|5| (9 standards + 3 samples) x (2 replicates) x (200 μL of WR) = 4800μL = 4.8mL WR (Prepare 7) 50 Reagent A:1 Reagent B 7mL:0.14mL

Notes: During the peperation step when nutrealizing the NaOH in the sample I used 280uL in the first sample but it was too acidic, I then added 20uL to the second by which it became a pH of 7. For the Third sample I added 20uL then another 20 after testing it then it became to acid reaching at a pH of 5

Test #3

All test samples were extras which were from Timepoint 4 |Samples| Airbrush Date|Tank| |——-|————–|—-| |DLextra| 20241016 |4 | |MD-4-27| 20241017 |5 | |DL-3-27| 20241025 |6 | |MD-1-36| 20241016 |3 | |DL-3-30| 20241017 |5 | |MD-2-35| 20241022 |2 | |DL-4-33| 20241022 |3 |

Added 10uL of NaOH; all samples had a pH of 10~ Samples at pH 10

(9 standards + 7 samples) x (2 replicates) x (200 μL of WR) = 6400μL = 6.4mL WR (Prepare 7) 50 Reagent A:1 Reagent B 7mL:0.128mL = 7000uL:128uL

#Chlorophyl a & c2 Protocol See HERE for the protocol used to attain chlorophyll concentrations All pratice samples were collected from Timepoint 4 |Samples| Airbrush Date|Tank|Box number placed in freezer| |DL-3-30| 20241017 |5 |6| |MD-1-29| 20241030 |
|DL-1-27| 20241031 | |6| |DL-2-28| 20241109 | |11| |MD-4-31| 20241106 | |11|

Changes made to the orginal protcol

• After thawing the sample votex for 15 seconds
• After centrifudge and adding 1mL of Acetone, vortex for more than 15seconds untill as much as the pellet that you possiblily can is disolved.