Shellfish Tissue Sampling For Genetic Analysis (Destructive)

GSI Protocol #05
Published: August 2023
Prepared by: Stephen G. Hesterberg, Ph.D., Executive Director; Sarah Hutchins, M.S., Staff Scientist

MATERIALS

• Ethanol (96% recommended, 95% acceptable)

• Shucking knife and gloves

• Dissection tools – snips, scalpel and tweezers

• Nitrile gloves

• Kim wipes and paper towels

• 2mL microcentrifuge tubes (sample <5 mm) or 5mL falcon tubes (sample >5 mm)

• Parafilm

• Pipettes

• Chemical labels and laboratory pen

• Shipping supplies – insulated box

INSTRUCTIONS

1. Collect a sufficient number of individuals to characterize the population (e.g., n = 30).

2. Use sterile techniques when working with multiple samples to prevent cross-contamination. Disinfect gloves, equipment, and work surfaces with ethanol between each sample.

3. Label the microcentrifuge or falcon tubes with chemical labels and a laboratory pen. The label should include a unique identifier (e.g., Site-Replicate-Date) and number of individuals per vial (e.g., n = X).

4. Individuals <5 mm shell length can be placed directly into a microcentrifuge or small falcon tube. Fill vial to the brim with ethanol.

5. Individuals >5 mm shell length should be dissected, using a scalpel or shucking knife to open valves. With dissecting tools, sample both adductor muscles and mantel and/or siphon tissues, blot tissue dry with paper towel, cut into small pieces, and place into vial (sample should be 10% of total volume). Fill vial to the brim with ethanol.

6. At the end of the sampling day, replace ethanol for improved fixation and seal vials with parafilm.

7. If shipping the same day, put sealed samples into an insulated box and overnight ship. If shipping is delayed, store samples in fridge (4°C) and replace ethanol at 24 h and 48 h.

REFERENCES

Protocol adapted from Dr. Huiping Yang and Dr. Xianyuan Zeng, Fisheries and Aquatic Science Program, School of Forest, Fisheries, and Geomatic Sciences, University of Florida.