The Rhodeus amarus (European bitterlingeDNA detection kit is able to detect the fish species Rhodeus amarus in all water samples worldwide and contains enough materials for 200 reactions. 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 Rhodeus amarus.
    The Rhodeus amarus qPCR detection kit has been validated for use in Dutch waters. Other sub species of Rhodeus amarus (R. colchicus, R. meridionalis, R. sericeus), which do not occur in Dutch waters, can also be detected with this primer set. Only use this kit if you are sure these additional species are not in your region or if you are interested in all these (sub)species.

    Species information

    The European bitterling (Rhodeus amarus) is a temperate freshwater fish belonging to the subfamily Acheilognathinae of the family Cyprinidae. It originates in Europe, ranging from the Rhone River basin in France to the Neva River in Russia. It is known simply as “the bitterling” in its native range, where it is the only species of its genus Rhodeus, and sometimes in the scientific literature, also, but this is technically wrong, being a leftover from the times when the European bitterling was united with its Siberian relative, the Amur bitterling, in R. sericeus. Properly, “bitterling” can refer to any species of Acheilognathus or Rhodeus.

    The fish reaches a size of up to 10 cm (4 in). It is found among plants over sandy and muddy bottoms in shallow waters. It feeds mainly on plants, and to a lesser extent, upon worms, crustaceans, and insect larvae.

    This species of fish was once used for human pregnancy tests. Female specimens were injected with the urine of the woman to be tested. If the woman was pregnant, the hormones in the urine would cause the fish’s ovipositors to protrude.

    The fish spawns in clear, slow-running or still water, often with a muddy bottom. The female deposits her eggs inside freshwater mussels. The male sheds his sperm into the inhalent current of the respiring mussel and thereby fertilizes the eggs. The young hatch and remain protected within the mussel for about a month, eventually leaving the mussel as actively swimming larvae. Both sexes reach sexual maturity in 1 year, at a length of 30 to 35 mm (1.2 to 1.4 in).

    In English: European bitterling)
    In German: Bitterling
    In French: Rhodeus amarus
    In Dutch: Bittervoorn

    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 200 reactions.

    • Positive control (Rhodeus amarus DNA)
    • 2x Sylphium qPCR mix (100 reactions) without primers and probes
    • 2x Primer/probe mix (100 reactions) for detection of Rhodeus amarus (FAM dye)
    • 1x Taq DNA polymerase (200 reactions)
    • Protocol and primer/probe validation report





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    Rhodeus amarus eDNA qPCR Detectiekit
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