Coral Bleaching Experiment Marcel Alcide Yao Djilan Leonard Desir Lauren Campbell

Procedure 1) Determine the Zoox density 2) Chlorophyll extraction & measurement 3) Determine protein concentration.

1) Determination of the zoox density Weighed 4 polyps 7.7 x 10-3g insertion of polyps into tube containing 2mL sea water. Homogenize for 20 sec. Measure the total homogenate volume/ 4.3 x 108 zoox/g

2) Chlorophyll extraction & measurement Addition of 9mL of acetone to 1mL of homogenate into new tube Vortex for 10 mn Centrifuge for 5mn Decantation Calculation of the amount of chlorophyll

Calculation of the amount of chlorophyll Wavelength 665nm → 647nm → 630nm → Absorbance 0.03 0.015 0.019

Ca = (11.85A665)-(1.54A647)-(0.08A630) 0.33088

Determination of the protein concentration .1mL of .25 mg/mL BSA + 5mL Bradford .1mL of .50mg/mL BSA + 5mL Bradford .1mL of .75 mg/mL BSA + 5mL Bradford .1mL of 1.00 mg/mL BSA + 5mL Bradford .1mL of 1.25 mg/mL BSA + 5mL Bradford .1 ml of homogenate+ 5mL of Bradford Abs(avg) .385 .641 .900 1.116 1.284 .074 Wavelength used: 595nm

Graph

Result .074 = 1.0139x + .0873 X = -.013

Discussion Why did we get a negative result? The use of an insufficient amount of polyps could cause that to happen because the machine used to measure the absorbance is very sensitive. In the next experiment we will use a greater amount of polyps to see if we will get a positive result.

The End