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Method for identifying Hucho taimen and Brachymystax lenok

A technology for salmonids and salmonids, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as mixed germplasm, achieve simple operation, improve work efficiency, and save money The effect of cost and time

Pending Publication Date: 2019-08-27
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The present invention provides an accurate method for distinguishing between salmon and lecherous salmon, which helps to solve the problems of germplasm mixing when seedlings are purchased and wild seedlings are collected, and also contributes to germplasm protection, rational utilization and germplasm biodiversity. Sex research provides technical support

Method used

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  • Method for identifying Hucho taimen and Brachymystax lenok

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Experimental program
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Effect test

specific Embodiment approach 1

[0012] Specific implementation mode one: the method for identifying the genetic sex of salmon in this implementation mode:

[0013] 1. Extract the genomic DNA of the sample to be identified;

[0014] 2. Perform PCR amplification using the genomic DNA in step 1 as a template. In the PCR amplification system, the upstream primer of the discrimination primer pair is 5'-ATCAATCATGGCTGGAGGTG-3', and the downstream primer of the discrimination primer pair is 5'-CACCCACACACGTACCAGTC-3';

[0015] 3. Gel electrophoresis of the PCR product. If the amplified band size is 133bp, it is called salmon, and if the amplified band size is 814bp, it is called salmon.

specific Embodiment approach 2

[0016] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the PCR amplification reaction system in step 2 is 20 μl: 2×Taq PCR mix 10 μl, the genomic DNA of the sample to be identified is 2 μl, the concentration of the discrimination primer is 10 μM, and the discrimination primer 1 μl each of the upstream and downstream primers of the pair, and the rest were supplemented with ultrapure water; the PCR reaction was: pre-denaturation at 95°C for 3 minutes, followed by 32 cycles of denaturation at 95°C for 30 sec, annealing at 66°C for 30 sec, and extension at 72°C for 30 sec; finally, extension at 72°C 5min. Others are the same as the first embodiment.

[0017] Under the PCR amplification reaction conditions of this embodiment, the target sequence is completely denatured without affecting the activity of the Taq enzyme, the primers are completely extended, an effective amplification amount can be achieved, and the non-specific amplification is small.

specific Embodiment approach 3

[0018] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is:

[0019] Step 1: Take 5-10 scales or 0.4cm 2 The fin rays were lysed with 40 μl lysate, and then digested with a PCR instrument. The digestion process was 55°C for 50 minutes and 98°C for 10 minutes; after that, mix well, and then centrifuge at 1000-2000rpm for 1-2 minutes, and take the supernatant to obtain the sample Genomic DNA. Others are the same as the first or second embodiment.

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Abstract

The invention discloses a method for identifying Hucho taimen and Brachymystax lenok, and relates to a fish identifying method. The method can be used for identifying a primer pair of the Hucho taimenand the Brachymystax lenok, wherein an upstream primer is 5'-CCAGGCGACATGGAGATG-3', and a downstream primer is 5'-CATCCGCGTCCCTTATGTAG-3'. The method can quickly and efficiently identify the Hucho taimen and the Brachymystax lenok by comparing the lengths of agarose gel electrophoresis amplification products thereof, and sequencing is not needed.

Description

technical field [0001] The invention relates to a fish identification method. Background technique [0002] Both Hucho taimen (Pallas) and Brachymystax lenok (Pallas) belong to the order Salmoniformes and the family Salmonidae. Closely related species, there are reports of natural hybrids between the two. Adult fish can be distinguished according to their external appearance, but the appearance of juvenile salmon is very similar to that of trout, and it is difficult to distinguish the two based on the external appearance alone, especially fish eggs, which are also difficult for professionals to distinguish. At present, breakthroughs have been made in the artificial propagation of jellyfish and leek salmon, and there are commercial seedlings for sale, but there is no report on the method for distinguishing jelly salmon and leek salmon juveniles. Contents of the invention [0003] The present invention provides an accurate method for distinguishing between salmon and leche...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6879C12N15/11
CPCC12Q1/6888C12Q1/6879
Inventor 匡友谊王荻唐国盘佟广香张庆渔徐伟尹家胜
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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