KIT PROTOCOL GREAT CRESTED NEWT eDNA KIT

For use in the UK only - for UK licencing

• Please ensure all information sent to NatureMetrics is in English.
  
• Datasheets must be completed using a pencil only.
• Kits should be used within three months.

^{Remember, always contact us to arrange sample collection. Click here to raise a logistics request with our customer support team.}

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Return to view all protocols

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IN THE FIELD

Gloves should be worn at all times during the sampling process. If you have purchased multiple kits, they should be used one at a time to minimize contamination between samples. Do not open more than one kit at a time. Samples should be collected from the shoreline without entering the water. This prevents disturbance of the substrate and limits cross-contamination.

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YOU WILL NEED

• Device to capture GPS coordinates (lat / long) (e.g. GPS capable smartphone or standalone GPS device)
• Pencil (to complete the datasheet)

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More than one person using the kit? Please purchase extra gloves.
To minimize waste we provide two pairs of gloves per kit. If two or more people will be handling the contents of the kit and collecting an eDNA sample, you will need to purchase additional pairs of gloves – enough for two pairs of gloves for each individual handling the kit. We recommend purchasing medical-grade nitrile gloves.

TIPS FOR GETTING THE BEST RESULTS

Avoiding Inconclusive Results:

To get the best results possible, to avoid inconclusive results, and to avoid situations where data cannot be reported from a sample, please read our HOW TO AVOID INCONCLUSIVE RESULTS guide before undertaking fieldwork.

In summary, when inconclusive results occur, it is because the target DNA was not detected, DNA was degraded, or PCR (a crucial part of the lab process) was inhibited.

Key steps you can take to help avoid inconclusive results: do not leave your samples exposed to sunlight or heat, and return samples to NatureMetrics as soon as possible after sampling.

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Avoiding contamination:

To increase confidence in results, avoiding contamination of samples is highly important. Sealed kits are sterile until opened; do not open more than one kit bag at a time, keep kit contents inside the bag before use, and put on the provided gloves before touching other kit components. Whilst wearing gloves, avoid touching anything not necessary to sample collection to minimize introducing external DNA into the sample. Change gloves between each kit. If any reusable tools are used, these should be decontaminated, using a disinfectant wipe or a bleach solution, between each sample.

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USER PROTOCOL

1. Before going into the field, ensure that you have confirmation of courier collection. If not, please book your sample return via the Customer Support Desk. Courier collection should be on the day of sample collection or as soon as possible after sample collection.

2. Identify where you will take 20 subsamples from the pond. Subsamples should be spread out evenly around the pond perimeter. If some of the pond perimeter is inaccessible, subsamples should be spread out evenly around accessible areas. Both open water (displaying habitat) and vegetated (egg-laying substrate) areas (if present) should be represented by the subsamples.

3. Put on one pair of provided gloves. Open the provided sampling bag by tearing off the clear plastic strip along the perforated line, then pulling the tabs to open the bag.

4. Collect 20 x 30 mL subsamples of pond water from around the pond (see step 2) using the provided ladle (fill the ladle to the top) and empty each subsample into the sampling bag. At the end, the sampling bag should be approximately half full (600 mL).

Before each ladle of water is taken, the pond water column should be gently mixed. Use the ladle to stir the water from the surface to close to the pond bottom without disturbing the sediment on the pond bottom. It is advisable not to sample very shallow water (less than 5-10 cm deep). The sampling bag is not self-standing so it should be propped against a rock/log/tree stump when it needs to be set down.

5. Once 20 subsamples have been taken, roll down the top of the sampling bag to seal, and shake the bag for 10 seconds. This mixes any DNA present in each subsample across the entire water sample.

6. Put on a new pair of gloves to minimize contamination risk to the next stage.

7. Before taking each aliquot, swirl the water in the sampling bag to homogenise the sample – this is because DNA will constantly sink to the bottom. Using the clear plastic pipette provided, take a 15 mL aliquot of water from the sampling bag and pipette into a sterile 50 mL conical tube containing 35 mL of ethanol and preservative (i.e. fill tube to the 50 mL mark). Close the tube, ensuring the cap is on tight. Shake the tube vigorously for 10 seconds to mix the water with the ethanol and preservative, then place it back into the cardboard insert.

8. Repeat step 7 for each of the 6 tubes in the kit. Seal the cardboard box containing the samples.

9. Scan the barcode on the box using the NatureMetrics app and submit the data for your samples. Complete and sign the sampling datasheet with a pencil (do not use pen as ink can easily run, to the point where it becomes unreadable). Please record your sampling data on paper and through the app so there is a physical and electronic copy of your data.

10. Place the cardboard box with samples and the sampling datasheet inside the resealable bag. Empty the remaining water in the sampling bag back into the pond.

11. The samples are collected and couriered to the NatureMetrics laboratory at ambient temperature. If samples are stored overnight before shipping, they should be refrigerated at 2-4°C. Samples can be stored for up to one month in the refrigerator before analysis, but should not be frozen or stored at ambient temperature.

12. Dispose of all waste responsibly. We accept back all waste from our sampling kits. All plastic waste must be well rinsed with tap water and dried prior to sending back. Any soiled, wet, or dirty plastic waste cannot be recycled and violates international permitting regulations, so washing and drying is essential. Plastic waste can be shipped back in the same package as the samples but must be packed in a separate bag to prevent any possible contamination. Please contact us via our customer support desk here, to return plastic waste free of charge.

Please ensure you have booked your courier collection by contacting our Logistics team via our customer support desk here, and booked your analysis on my.naturemetrics.co.uk.

Contact our GCN team for support at gcn@naturemetrics.co.uk.

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PACKAGING NOTES AND QUICK LINKS

• Return your samples using the box that kits were sent in, where possible.
• If using an alternative box, ensure that it is in good condition and remove any old labels.
• Avoid excess spacing by adding cushioning material (e.g. bubble wrap, newspaper) to avoid the product shifting inside the box.
• Use good quality sealing tape and seal along box edges in a H pattern.
• The only essential thing to send back to us is the specimen bag containing the filter and the sampling datasheet. However, all kit contents can be sent back to NatureMetrics for recycling.
• Please find our Terms and Conditions here.

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CLIENT RESPONSIBILITIES

Important considerations for your sampling

• Samples from some locations might have compounds that inhibit analyses resulting in reduced data generation – while we routinely test for inhibition, it is not always possible to overcome.
• It is the responsibility of the client to ensure that all efforts have been made to avoid contamination from external sources and between samples.
• Handling samples without gloves can increase the content of human DNA, reducing data generation.
• After sampling, movement of samples across borders without permission is not allowed as they are classed as biological samples.

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Disclaimer: Safe sample collection is the responsibility of the Client. NatureMetrics accepts no liability associated with the use of the kits and sample collection. The Client is solely responsible for the quality of the samples and the representativeness of the samples received by NatureMetrics. Please read the full limitation of liability statement in the Terms and Conditions.

See the following advice note for sampling tips, as well as information on control samples and avoiding contamination.

View Advice Note

Health & Safety information for this kit can be viewed here.

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