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This method for separating coral tissues from algal endosymbiont (Symbiodiniaceae) for stable isotope analysis is modified from previously published methods (Hughes et al. 2010). There are three parts to preparing coral samples for stable carbon and nitrogen isotope analysis: 1) airbrush to remove coral tissue and algal cells from skeleton and store at -80 °C until ready to separate, 2) separate the coral tissue from the algal cells through centrifugation and filtering, and 3) dry and pack separated tissues into tin capsules for analysis in a stable isotope ratio mass spectrometer. This method was modified from Hughes et al. (2010) by James Price with the assistance of Alex Smith and Kerri Dobson and with the guidance of Andréa Grottoli at The Ohio State University. dx.doi.org/10.17504/protocols.io.bgi7juhn 

This protocol outlines a method for quantifying the total biomass of Scleractinian coral samples which have been ground into a homogenous paste consisting of aragonite skeleton, coral host tissue, and endosymbiotic Symbiodiniaceae cells. There are four parts to quantifying total biomass: 1) grind coral fragments into a homogenous paste, 2) partition the biomass subsample, 3) quantify the ash-free dry weight [AFDW], and 4) standardize AFDW to the colony surface area. This method has been reported in several publications by Grottoli's team (e.g., Rodrigues & Grottoli 2007). This protocol was written by Rowan McLachlan (03-19-20) and was reviewed by all co-authors. dx.doi.org/10.17504/protocols.io.bdyai7se 

This method for extracting protein from ground coral samples is based on the Bradford assay for the colorimetric detection and quantification of total protein (Bradford, 1976) and is compared to a known standard dilution of bovine serum albumin (BSA). Pierce Inc. and Bio-Rad have developed the reagents and standards necessary for completing the extraction. There are five parts to quantifying total soluble protein in ground corals: 1) grind and sub-sample the coral and store at -80 °C until ready to extract, 2) solubilize protein via cell disruption [detergent lysis and freeze-thaw lysis], 3) separate the dissolved protein from tissue and skeletal particles, 4) quantify the protein concentration via Bradford microassay procedure, and 5) standardize the protein concentration to ashfree dry weight (AFDW). This method was originally developed by Rowan McLachlan with the assistance of Jamie Price and Kerri Dobson and with the guidance of Dr. Noah Weisleder and Andréa Grottoli at The Ohio State University. This protocol was written by Rowan McLachlan and reviewed by all co-authors. dx.doi.org/10.17504/protocols.io.bdc8i2zw 

Abstract Coral bleaching is the single largest global threat to coral reefs worldwide. Integrating the diverse body of work on coral bleaching is critical to understanding and combating this global problem. Yet investigating the drivers, patterns, and processes of coral bleaching poses a major challenge. A recent review of published experiments revealed a wide range of experimental variables used across studies. Such a wide range of approaches enhances discovery, but without full transparency in the experimental and analytical methods used, can also make comparisons among studies challenging. To increase comparability but not stifle innovation, we propose a common framework for coral bleaching experiments that includes consideration of coral provenance, experimental conditions, and husbandry. For example, reporting the number of genets used, collection site conditions, the experimental temperature offset(s) from the maximum monthly mean (MMM) of the collection site, experimental light conditions, flow, and the feeding regime will greatly facilitate comparability across studies. Similarly, quantifying common response variables of endosymbiont (Symbiodiniaceae) and holobiont phenotypes (i.e., color, chlorophyll, endosymbiont cell density, mortality, and skeletal growth) could further facilitate cross‐study comparisons. While no single bleaching experiment can provide the data necessary to determine global coral responses of all corals to current and future ocean warming, linking studies through a common framework as outlined here, would help increase comparability among experiments, facilitate synthetic insights into the causes and underlying mechanisms of coral bleaching, and reveal unique bleaching responses among genets, species, and regions. Such a collaborative framework that fosters transparency in methods used would strengthen comparisons among studies that can help inform coral reef management and facilitate conservation strategies to mitigate coral bleaching worldwide.