The Pelobatus fuscus (Common spadefoot) eDNA detection kit is able to detect the amphibian species Pelobatus fuscus in all waters worldwide and contains enough materials for 20 samples to be analysed in 8 fold (200 qPCR tests). The kit is based on the fast, sensitive, and proven primers/probe qPCR technique. The kit contains primers and a probe for detecting a highly specific sequence present on the mitochondrial DNA of the species Pelobatus fuscus.
Species information
Common spadefoot (Pelobates fuscus) is a species of toad in the family Pelobatidae, native to an area extending from Central Europe to Western Asia. It is commonly known as the common spadefoot, garlic toad, the common spadefoot toad and the European common spadefoot.
The common spadefoot grows to a length of approximately 6.5 centimetres (2.6 in) for males and 8 centimetres (3.1 in) for females. The skin colouration varies depending upon habitat, gender and region, but is usually light-grey to beige-brown on the dorsal surface. The skin is mottled by darker marks that differ between individuals. The belly is white, sometimes with grey mottling. Albino specimens have been observed. When alarmed, it emits a very loud call (alarm call) and it can exude a noxious secretion which smells like garlic, hence the common name “garlic toad”.
In English: common spadefoot, garlic toad, the common spadefoot toad and the European common spadefoot.
In German: Knoblauchkröte.
In French: Pélobate bruni.
In Dutch: Knoflookpad.
Primer design
The primers and probe are specially designed to be used with eDNA samples and have the following properties:
- Highest possible sensitivity (1 DNA copies per reaction). Environmental water samples contain normally very low amounts of target DNA.
- Strong fluorescence signal with low background noise. Isolated environmental samples contain residues of naturally occurring auto fluoresce substances that will interfere with the measurements. A strong fluorescence signal from the analyses is required for these kind of samples.
- 100% specificity. Isolated DNA from environmental samples contains billions of DNA fragments from bacteria, protozoa, plants, animals, etc.
The kit is developed and optimized to be used on eDNA isolates purified using the eDNA isolation kit (#SYL002) from Sylphium molecular ecology.
Kit contents
The kit contains materials for an 8 fold analyses on 20 samples including all necessary controls.
- Positive control (Cloned Pelobates fuscus DNA)
- 2x Sylphium qPCR mix (100 reactions) without primers and probes
- 2x Primer/probe mix (100 reactions) for detection of Pelobates fuscus (FAM dye)
- 1x Taq DNA polymerase (200 reactions)
- Protocol and primer/probe validation report
Manual
