Description
The Pseudorasbora parva (stone moroko) eDNA detection kit is able to detect the fish species Pseudorasbora parva 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 Rana arvalis.
Detection as a service
We offer the detection of Pseudorasbora parva also as a service. See our eDNA service website (in Dutch) or contact us for more information about sampling, sending, and pricing.
Species information
The stone moroko (Pseudorasbora parva), also known as the topmouth gudgeon, is a fish belonging to the Cyprinid family, native to Asia, but introduced and now considered an invasive species in Europe. The fish’s size is rarely above 8 cm and usually 2 to 7.5 cm long.
The fish was introduced in the 1960s into ponds in Nucet, Dâmboviţa County, Romania and it made its way into Danube, then spreading throughout Europe. They pose danger to other species such as the sunbleaks (Leucaspius delineatus). They are the carrier of a parasite (Sphaerothecum destruens) that is not damaging to the topmouth gudgeon, but attacks other fishes like the sunbleaks, which are unable to spawn and have a higher mortality when infected.[4] They also feed on eggs of locally valuable native fish species.
The species has also been recently discovered in several lakes in the UK where it is believed to have been illegally stocked. This has called for a large scale eradication programme organised by the Environment Agency who kill the fish off with a piscicide called rotenone. (source: wikipedia)
In English: stone moroko or topmouth gudgeon.
In German: Blaubandbärbling.
In French:
In Dutch: Blauwband.
Primer design
The primers and probe are specially designed to be used with eDNA samples and have the following properties:
- Highest possible sensitivity (<10 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. Not only animals from the same order (fish) must be taken in account during primer/probe design, but all known DNA sequences must be checked for nonspecific binding of the primers and probe. This is validated by experimental and bioinformatical studies.
The kit is developed and optimized to be used on eDNA isolates purified using the eDNA isolation kit (#SYL002) from Sylphium molecular ecology.
Other eDNA isolation methods/kits can be used as well. Please contact us to get more information how to use the obtained isolates from other methods/kits to get reliable results with the Pseudorasbora parva eDNA detection kit.
Kit contents
The kit contains materials for an 8 fold analyses on 20 samples including all necessary controls.
- Positive control (Pseudorasbora parva DNA)
- 2x Sylphium qPCR mix (100 reactions) without primers and probes
- 2x Primer/probe mix (100 reactions) for detection of Pseudorasbora parva (FAM dye)
- 1x Taq DNA polymerase (200 reactions)
- Protocol and primer/probe validation report
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