INTRO

Families 5 & 7 from the June 2026 SORMI experiment (GitHub repo) were selected for citrate synthase assay (selection by Steven). Ctenidia samples from these families were homogenized in preparation for the Citrate Synthase Assay Kit (ABCAM; cat: ab239712).

MATERIALS & METHODS

Eight ctenidia samples (1-8) from each treatment (ambient and 36°C) in Families 5 and 7 were selected. Samples were stored on dried ice until homogenization. Pieces of ctenidia were broken off and weighed to the nearest 0.1 mg. Tissue was transferred to 1.5mL Safe-Lock tubes (Eppendorf) containing 350 µL of Assay Buffer 7 and ~100µL of 0.5mm glass beads (Research Products International). Samples were homogenized in a pre-/cooled (dry ice) Bullet Blender 5E Gold+ using the 1.5mL adapters. Homogenization was performed for 10 minutes at Speed 12. After homogenization, samples were stored at -80°C until further processing.

Note

A homogenization volume of 350 µL (instead of the manufacturer’s recommended 100 µL) was used based off of prior work in our lab with this kit, performed by Matt George (link to the paper - requires institutional access).

Below is a list of the samples and the weights of ctenidia used for homogenization:

Sample	Weight(mg)
F05_01_ambient	10.1
F05_02_ambient	17.5
F05_03_ambient	22.3
F05_04_ambient	14
F05_05_ambient	11.5
F05_06_ambient	30.2
F05_07_ambient	25.8
F05_08_ambient	30.6
F05_01_36C	12.8
F05_02_36C	6.2
F05_03_36C	10.3
F05_04_36C	12.8
F05_05_36C	22.7
F05_06_36C	18.4
F05_07_36C	11.3
F05_08_36C	13.8
F07_01_ambient	7.3
F07_02_ambient	19.4
F07_03_ambient	19.7
F07_04_ambient	10.7
F07_05_ambient	8.3
F07_06_ambient	12.8
F07_07_ambient	20.6
F07_08_ambient	10.9
F07_01_36C	9
F07_02_36C	9.5
F07_03_36C	18.3
F07_04_36C	16.9
F07_05_36C	10.5
F07_06_36C	9.7
F07_07_36C	12.5
F07_08_36C	9.1

20240811-mgig-Fams5_and_7-citrate_synthase-tissue-weights (Google Sheet)