Water laboratories and ecological consultancies can now perform DNA analyzes themselves in their own (to be set up) laboratory by using kits developed by Sylphium molecular ecology. Doing it yourself has the advantage that you can schedule the research yourself at the time you want and that the results are available faster. An additional advantage is that you have full control over the entire process from sampling to reporting. Performing the analyses yourself also leads to a large cost reduction of eDNA determinations.
How does it work?
Sylphium molecular ecology has developed various kits to perform the most sensitive and reliable eDNA determinations with as few actions as possible. The entire process from sampling, extraction of eDNA up to the qPCR analysis can be performed using these kits. All work with these kits should be performed in a laboratory specially set up for eDNA work. Sylphium gives full support in setting up an eDNA lab. If you are interested in these courses and support, please contact us.
Setup of an eDNA laboratory
Contaminations with DNA can be a problem due to the high sensitivity of the qPCR methods. Positive controls and amplifications products after PCR (post-PCR) contain large amounts of specific DNA fragments and are therefore important potential sources of contamination. It is important to organise the laboratory activities in such a way that the risk of contamination is minimised.
To minimise the chance of contamination when using PCR, the advice is to use two physically separated rooms:
Reagent preparation room
The reagent preparation room is used for the preparation and storage of PCR reagents. In this room, the PCR mix is assembled and divided over the reaction vials of the PCR plate. After filling, the plate will be covered and transported to the sample preparation room.
Sample preparation room
The sample preparation room is intended for extraction of DNA from samples and positive and negative controls. The extracted DNA will also be added to the PCR mix in this room. Care must be taken while adding the positive control to the mix. A PCR cabinet with a UV lamp can be used to prevent spreading of the positive control. The PCR vials are sealed once the DNA has been added and transported to the PCR equipment, which is preferably placed outside the lab.
Although two separate rooms is preferable, eDNA extraction and preparation of PCR reagents can also performed in a single room. Therfor two PCR cabinets with a UV lamps can be used instead. These cabinets can be seen as separate rooms. In this case, space is limited.
Furthermore, specific training courses are provided to properly implement the various eDNA technologies and full implementation of the various techniques will be ensured.
- UV work cabinet(s) with UV light
- qPCR device
- Centrifuges for 2 ml and 13 ml tubes
- pipette sets (2x)