Molecular marking primer and method for identifying Chinese coastal platycephalus indicus species indeterminate (sp) and platycephalus cultellatus

A technology of molecular markers and molecular marker sequences, which is applied in the field of molecular marker primers for identifying unspecified species of flatfish sp and flatfish along the coast of China, and achieves the effects of simple identification method, appropriate length, and consistent amplification results

CN107557483AInactive Publication Date: 2018-01-09ZHEJIANG OCEAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2018-01-09
Estimated Expiration
Not applicable · inactive patent
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Abstract

The invention discloses a molecular marking primer and method for identifying Chinese coastal platycephalus indicus species indeterminate (sp) and platycephalus cultellatus. The sequence of the molecular marking primer Fish F1 is 5'-TCAACCAACCACAAAGACATTGGCAC-3', and the sequence of the molecular marking primer Fish R2 is 5'-ACTTCAGGGTGACCGAAGAATCAGAA-3'. The molecular marking method includes: extracting DNA of platycephalus indicus species indeterminate (sp) and platycephalus cultellatus, and taking the DNA of the two fish as templates for PCR amplification, performing purification and sequencing to the amplification results, and comparing the sequencing results. The molecular marking primer is proper in length, is closely complementary to the sequences of the templates, and is beneficialfor an amplification reaction. The molecular marking method is simple, the identification result is stable, and the repeatability is good. The amplification result of Chinese coastal platycephalus indicus species indeterminate (sp) is consistent, so that the primer and method show high consistency.
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Description

technical field

[0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker primer and a method for identifying undetermined species of flatfish sp and flatfish along the coast of China. Background technique

[0002] The undetermined species of flatfish, sp, belongs to the order Sculliformes, the suborder, the family, and the genus Flatfish. It is only distributed along the coast of the South China Sea in China. Its external shape is very similar to that of swordfish: dorsal fin rays II-VII-I+13; anal fin rays 13; pectoral fin rays 18-20 (mainly 18 or 19); pelvic fin rays I, 5; caudal fin forked rays 9-12 (mainly 12). Lateral line scales 67-76; body shorter, flattened, tapering backwards. Kiss flat, slightly elongated. Mouth large, superior, extending back beyond the front edge of the eyes. Eyes small, no depression behind eyes. The interopercle has no appendages. The lower edge is smooth, with only 1 spine....

Examples

Embodiment 1

[0018] A molecular marker primer for differentiating the undetermined species of flatfish sp from the coast of China, and the genus sp., the primer sequence is:

[0019] Fish F1:5'-TCAACCAACCACAAAGACATTGGCAC-3';

[0020] Fish R2: 5'-ACTTCAGGGTGACCGAAGAATCAGAA-3'. The above-mentioned primers are of appropriate length and are closely complementary to the sequence of the template. It is difficult to form a stable dimer and hairpin structure between the primers, and the priming efficiency at the mismatch site is extremely low. There is no complementary sequence by itself, and it is difficult to form by itself. The hairpin structure can be closely complementary to the sequence of the template, and the extension temperature is less than 74°C, which is beneficial to the amplification reaction.

[0021] A molecular marker method for identifying the undetermined species of flatfish sp from the coasts of China and the common species of flatfish. The specific steps are as follows: Selec...