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Primer group, kit and method for identifying tachysurus fulvidraco and leiocassis longirostris hybrid species

A technology for yellow catfish and long-nose catfish, which is applied in the field of primer sets of male and female parents, and can solve problems such as methods without offspring

Inactive Publication Date: 2016-06-22
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hybridization of Pelteobagrus fulvidraco and Leiocassis longirostris has begun, and their offspring have shown good growth advantages, but so far there is still no method for identifying their offspring. The method of hybrid individual identification is especially necessary

Method used

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  • Primer group, kit and method for identifying tachysurus fulvidraco and leiocassis longirostris hybrid species
  • Primer group, kit and method for identifying tachysurus fulvidraco and leiocassis longirostris hybrid species
  • Primer group, kit and method for identifying tachysurus fulvidraco and leiocassis longirostris hybrid species

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 is used to identify the primer set of the male parent and the female parent of the yellow catfish and the long-nose catfish hybrid

[0030] The present invention is used to identify the male parent and the female parent of the yellow catfish and the long snout hybrid. The primer set consists of two sets of primer pairs, and these two sets of primer pairs are respectively:

[0031] A pair of primers for amplifying the COI gene, wherein the forward primer COIF is the sequence shown in SEQ ID NO.1: 5'-CTACAATCCACCGCCTAA-3', and the reverse primer COIR is the sequence shown in SEQ ID NO.2: 5'-TAGAAGAAAGTGACAGAGCG- 3';

[0032] A pair of primers for amplifying the ITS sequence, wherein the forward primer ITSF is the sequence shown in SEQ ID NO.3: 5'-CGTAACAAGGTTTCCGTAGGTG-3', and the reverse primer ITSR is the sequence shown in SEQ ID NO.4: 5'-ATCCACCGCTAAGAGTTGTCAG- 3'.

Embodiment 2

[0033] Example 2 is used to identify the kit of male and female parents of yellow catfish and long snout hybrids

[0034] A kit invented for identifying hybrids of yellow catfish and long-nosed catfish contains the following components:

[0035] 1) A pair of primers for amplifying the COI gene, wherein the forward primer COIF is the sequence shown in SEQ ID NO.1: 5'-CTACAATCCACCGCCTAA-3', and the reverse primer COIR is the sequence shown in SEQ ID NO.2: 5'- TAGAAGAAAGTGACAGAGCG-3';

[0036] 2) A pair of primers for amplifying the ITS sequence, wherein the forward primer ITSF is the sequence shown in SEQ ID NO.3: 5'-CGTAACAAGGTTTCCGTAGGTG-3', and the reverse primer ITSR is the sequence shown in SEQ ID NO.4: 5'- ATCCACCGCTAAGAGTTGTCAG-3';

[0037] 3) 10× buffer for PCR;

[0038] 4) dNTPs;

[0039] 5) Taq DNA polymerase;

[0040] 6) Cloning vector pMD18-T;

[0041] 7) Solution (including T4DNA ligase and its buffer);

[0042] 8) LATaq DNA polymerase;

[0043] 9) 2×GC buffer...

Embodiment 3

[0045] Example 3 Using the above-mentioned kit to identify the male and female parents of the hybrids of Pelida catfish and long-nose catfish

[0046] The method for identifying yellow catfish and long-nose catfish hybrids of the present invention comprises the following steps:

[0047] 1. Collect the caudal fins of 6 individuals S1-S6 to be identified, purebred yellow catfish H1-H3 and purebred long-nose catfish C1-C3, store them in absolute ethanol, and bring them back to the laboratory.

[0048] 2. Genomic DNA extraction

[0049] DNA was extracted using a Genomic DNA Extraction Kit (TIANGEN).

[0050] 3. Identification of the female parent of the hybrid

[0051] 3.1 COI gene PCR amplification

[0052] Using the DNA extracted as described above as a template, use the primer pair COIF / COIR to amplify the COI gene by PCR. The PCR amplification system is as follows:

[0053]

[0054] The PCR reaction process was: pre-denaturation at 94°C for 5 min, 35 cycles (denaturatio...

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Abstract

The invention provides a primer group for identifying a male parent and a female parent of tachysurus fulvidraco and leiocassis longirostris hybrid species. The primer group comprises a primer pair for amplifying COI genes and a primer pair for amplifying an ITS sequence, wherein the primer pair for amplifying COI genes consists of a primer COIF and a primer COIR; the primer pair for amplifying an ITS sequence consists of a primer ITSF and a primer ITSR. The invention also provides a kit which is for identifying the male parent and the female parent of tachysurus fulvidraco and leiocassis longirostris hybrid species and comprises the primer group. The kit also comprises any one or more of the following reagents: a buffer solution for PCR, dNTP, heat-resistant DNA polymerase, a cloning vector, a DNA ligase, and a buffer solution for ligation. The invention also provides a method for identifying the male parent and the female parent of tachysurus fulvidraco and leiocassis longirostris hybrid species. The primer group, the kit or the method can be used for accurately identifying the male parent and the female parent of the tachysurus fulvidraco and leiocassis longirostris hybrid species, thus effectively managing a breeding plan and monitoring negative influence thereof, and has great significance in sustainable development of aquaculture industry.

Description

technical field [0001] The invention relates to the field of aquaculture, and more particularly relates to a primer set, a kit and a method for identifying male and female parents of hybrids of yellow catfish and long-nose catfish. Background technique [0002] Hybrid breeding is an important method of fish breeding, which is to use species, strains or individuals with excellent traits to hybridize to produce new excellent traits, so that the offspring can be genetically improved, and heterosis can be used. In my country, we have successfully bred Feng carp, Heyuan carp, Yue carp, Furong carp, Oni tilapia, hybrid snakehead, Ganchang carp and crucian carp, hybrid carp "Pioneer No. 1", and hybrid bream through hybrid breeding technology. A large number of new varieties of aquatic hybrids have played an important role in aquaculture. [0003] Fishes of the family Oleopsis are widely distributed in the Yangtze River, Pearl River, Heilongjiang and Yellow River basins in my countr...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/158
Inventor 梁宏伟郭闪闪罗相忠李忠邹桂伟
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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