Putnam Lab Ash-Free Dry Weight Protocol

Draft Protocol

Protocol written for Montipora capitata and Pocillopora acuta adult coral samples from the Holobiont Integration 2018 project.

1. Turn on oven and set to 75°C.
2. Sonicate or homogenize tissue sample on cold PBS. For Holobiont Integration project, this step is done during the airbrushing protocol. The following steps are done with the Total Protein/Total Antioxidant Capacity aliquot.
3. Label a tin foil cup with the coral ID.
4. Weigh the empty tin foil cup. Record value (g) in Ash-Free Dry Weight spreadsheet.
5. Pipette X μL of coral tissue slurry into the tin foil.
6. Place the tin foil cup in the oven and heat for 24 hours.
7. Weigh the tin foil cup after 24 hours in the oven and record the value (g).
8. Place the tin foil cup in the muffle furnace for X hours at 450°C.
9. Weigh the tin foil cup after X hours in the muffle furnace and record the value (g).

Discard the tin foil cup in the garbage.