Universal primers and the use thereof for the detection and identification of amphibia/fish species
AssigneesUniversite Grenoble Alpes, Centre National De La Recherche Scientifique
InventorsPierre Taberlet, Eric Coissac, Tiayyba Riaz
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Abstract
The invention relates to oligonucleotides and to the use thereof as universal primers for the detection and identification of amphibia/fish species, especially in complex and deteriorated substrates. The invention also relates to a method for detecting and identifying amphibia/fish species in samples collected from the environment (ground, water, faeces) or from industry (especially transformed products). The invention further relates to a kit for said detection.
Claims9
A method for amplifying a region of the mitochondrial 12S gene of batrachian and fish species, comprising the following steps:
- a) a sample suspected of containing DNA from a batrachian and/or fish species is provided, the batrachian species is a part of the batrachian kingdom of amphibia;
- b) an amplification chain reaction is carried out using a pair of oligonucleotides, according to which the first oligonucleotide selectively hybridizes to the sequence SEQ NO. 4 and the second oligonucleotide selectively hybridizes to the sequence SEQ ID NO. 5 or to the sequence SEQ ID NO. 6.
A method for detecting a batrachian and/or fish species in a sample, comprising the following steps:
- a) a sample suspected of containing DNA from a batrachian and/or fish species is provided, the batrachian species is a part of the batrachian kingdom of amphibia;
- b) the total DNA contained in the sample is isolated;
- c) an amplification chain reaction is carried out using a pair of oligonucleotides, according to which the first oligonucleotide selectively hybridizes to the sequence SEQ NO. 4 and the second oligonucleotide selectively hybridizes to the sequence SEQ ID NO. 5 or to the sequence SEQ ID NO. 6;
- and
- d) the possible presence of an amplification product is detected.
A process for detecting and identifying a batrachian and/or fish species in a sample, comprising the following steps:
- a) a sample suspected of containing DNA from a batrachian and/or fish species is provided, the batrachian species is a part of the batrachian kingdom of amphibia;
- b) the total DNA contained in the sample is isolated;
- c) an amplification chain reaction is carried out using a pair of oligonucleotides, according to which the first oligonucleotide selectively hybridizes to the sequence SEQ NO. 4 and the second oligonucleotide selectively hybridizes to the sequence SEQ ID NO. 5 or to the sequence SEQ ID NO. 6;
- d) the presence of an amplification product is detected;
- and
- e) the sequence of the amplification product is determined in order to identify the batrachian and/or fish species contained in the sample.
Dependencies are inferred from the claim text.