Freshwater Habitats Trust How to Videos - eDNA kits

TL;DR

Key insights

eDNA Sampling for Great Crested Newts is a Key Component of PondNet Project

• The Freshwater Habitats Trust uses eDNA kits to monitor Great Crested Newts.
• This sampling is part of the 'People Ponds and Water PondNet' project.
• Each pond survey requires both an eDNA water sample and a pond habitat survey form.
• The entire process for both samples is estimated to take about an hour.

Proper Kit Usage and Contamination Prevention are Crucial for Accurate eDNA Results

• Each kit contains sterile gloves, a ladle, a well pack bag, a pet, and tubes with preservative fluid.
• A fresh kit must be used for every pond to avoid cross-contamination.
• Accurate labeling of the kit and form with identical information is essential for lab tracking.
• Sampling should be done in pairs for safety and to assist with bag handling.

Systematic Water Sampling Technique Ensures Comprehensive DNA Capture

• Twenty sampling locations should be identified and spread evenly around the pond.
• Sampling should include both open water and vegetated areas, avoiding shallow water (<10 cm) to prevent sediment contamination.
• The water column should be gently mixed from surface to near bottom without disturbing sediment.
• After collecting 20 samples into the well pack bag, it must be shaken for 10 seconds to mix the DNA.

Post-Sampling Procedures and Sample Handling are Vital for Preservation

• Water from the well pack bag is pipetted into sterile tubes containing preservative fluid, filling to the 50ml mark.
• Each tube must be securely capped and shaken for 10 seconds to mix DNA with the preservative.
• All six tubes should be filled, with the well pack bag shaken each time to ensure DNA doesn't sink.
• Collected samples should be stored in a cool place, out of direct sunlight, until collection or drop-off.

Key moments

• 00:00Introduction to the eDNA kit for surveying Great Crested Newts as part of the PondNet project, outlining the need for both eDNA samples and habitat survey forms.
• 00:30Inventory of the eDNA kit contents: sterile gloves, ladle, well pack bag, pet, and preservative tubes, stressing the use of a fresh kit per pond.
• 00:56Importance of completing the paperwork first, including names, pond name, grid reference, date, and the kit number, and transferring this information to the sample kit label.
• 01:31Procedure for collecting the eDNA sample: identifying 20 locations around the pond, spreading them evenly, avoiding shallow water and sediment, and ensuring safety by not entering the water.
• 02:11Step-by-step guide to collecting the water sample using the ladle and well pack bag, including how to open the bag and manage difficult access points with an extended ladle.
• 03:34Transferring the collected water samples into the sterile tubes with preservative fluid, filling to the 50ml mark, and shaking each tube for 10 seconds.
• 04:26Post-sampling instructions: emptying excess water back into the pond, keeping the sample box upright, and storing samples in a cool place before collection or drop-off.