CN202808812U

CN202808812U - Rapid duck gender identification kit body

Info

Publication number: CN202808812U
Application number: CN2012202861143U
Authority: CN
Inventors: 胡艳; 李慧芳; 顾华兵; 徐文娟; 章明; 宋迟; 陶志云; 单艳菊; 章双杰; 陈文峰
Original assignee: Jiangsu Tengdayuan Agriculture And Livestock Co Ltd; Jiangsu Institute Poultry Sciences
Current assignee: Jiangsu Tengdayuan Agriculture And Livestock Co Ltd; Jiangsu Institute Poultry Sciences
Priority date: 2012-06-18
Filing date: 2012-06-18
Publication date: 2013-03-20
Anticipated expiration: 2022-06-18

Abstract

The utility model discloses a rapid duck gender identification kit body, belonging to the field of biotechnologies. The rapid duck gender identification kit body consists of a kit body, a liner of the kit body, and a specific primer mixture ZWp, an amplification buffer solution, a DNA marker and agarose which are respectively arranged in each hole cavity of the liner. According to the kit body, a PCR (Polymerase Chain Reaction)-agarose electrophoresis technology is adopted to detect the fragment sizes of PCR products, and the gender of an individual is identified according to the fragment sizes and quantity of the PCR products. The rapid duck gender identification kit body has the advantages of high accuracy, rapidness and low cost.

Description

The quick identification reagent box of duck sex box body

Technical field

The utility model belongs to biological technical field, and a kind of test kit that carries out fast the sex identification of duck is provided, and this test kit is used for PCR-agarose electrophoresis technology.

Background technology

The sex of poultry is determined by W karyomit(e) and Z chromosome, poultry female gene type is the ZW type, and male genotype is the ZZ type, and the probability that Z or W pass to filial generation is the same, be 1/2nd, its mode of inheritance is followed the Mendelian law of independent assortment and the law of independent assortment.Up to the present, find to have minority to be distributed in gene on bird Z and the W karyomit(e), they do not appear at false euchromosome district, but all are to occur with W and the single copy of Z, and exist with the form of the non-restructuring part of two types in sex chromosome.

That have characteristic feature most is CHD1Z--CHD1W(chromodomain helicase DNA binding protein 1) gene, research thinks that their in all birds (except ratites) are chain with W or Z respectively.Because some introns are different from sequence size among the CHD1W at gene C HDIZ, use the conservative property primer, just can pass through PCR-agarose electrophoresis technology, clip size and quantity according to the PCR product, judge male genotype (ZZ type, a large PCR product fragment) and female gene type (ZW type, small one and large one two PCR product fragments).These two kinds of genes are independently evolved, and use the Direct PCR technical Analysis, for research birds (except ratite) molecule sex determination provides approach.Along with going deep into that the subjects such as fetology, developmental biology and molecular biology of duck are studied, the evaluation of duck embryonic early sex becomes a problem in science.Through the retrieval to existing domestic and foreign literature and patent, so far there are no the research report of the duck CHD of family gene complete sequence and molecular sex identification.

The utility model content

The purpose of this utility model is in order to provide a kind of rational in infrastructure, cheap gene type test kit to duck sex Molecular Identification.

The purpose of this utility model is achieved through the following technical solutions, the quick identification reagent box of a kind of duck sex box body, described test kit is by box body, liner in the box body, the Auele Specific Primer mixture ZWp that inserts respectively in each vestibule of liner, amplification buffer, DNA marker and agarose form.

Wherein the ZWp details are as follows:

Primer mixture ZWp represents that primer information is as follows for the upstream and downstream primer mixture of CHD gene PCR amplification:

Upstream primer 5'-AGTGCATTGCAGAAGCAATATT-3',

Downstream primer 5'-GCCTCCTGTTTATTATAGAATTCAT-3'.

This test kit using method:

1, pcr amplification purpose fragment

1) the pcr amplification reaction system is prepared

In the PCR thin-walled tube of 200 μ l, add 2 μ l template DNAs (about 50 ng), 12.5 μ l amplification buffers, 2 μ l primer ZWP (5uM) and 8.5 μ l water, the reaction system of PCR is 25 μ l, and is fully of short duration centrifugal behind the mixing.

2) pcr amplification reaction programming

The reaction conditions that PCR is set at eppendorf PCR instrument is: 94 ℃ of denaturation 4 min, and rear 94 ℃ of sex change 30s, 55 ℃ of renaturation 30s, 72 ℃ are extended 30s, 35 circulations, last 72 ℃ are extended 6 min, and 4 ℃ save backup.After programming is complete, the PCR reaction tubes is put into the PCR instrument react amplification.

2, identified gene type

Reaction is got 4-10 μ l reaction product at 2% agarose gel, electrophoresis in 1 * TBE electrophoresis liquid after finishing.Under the room temperature, according to electrophoresis chamber width * 6-8V, electrophoresis 20 minutes.According to electrophoretogram, the identified gene type.

The utility model is comprised of upstream and downstream primer mixture ZWp, amplification buffer, DNA marker and the agarose of CHD gene PCR amplification, utilizes the genotype of PCR-agarose electrophoresis technology for detection gene, has accuracy height, quick, cheap advantage.

Description of drawings

Fig. 1 is the structural representation of the quick identification reagent box of the utility model duck sex;

Fig. 2 CHD gene PCR product somatotype figure;

Among the figure: 1 box body, 2 liners, 3 Auele Specific Primer mixture ZWp, 4 amplification buffers, 5DNA marker, 6 agaroses.

Embodiment

Be described further the utility model with accompanying drawing in conjunction with the embodiments.Should be understood that these embodiment only are used for illustration purpose, and be not used in the restriction scope of the invention.

The quick identification reagent box of a kind of duck sex, by box body 1, the liner 2 in the box body, the Auele Specific Primer mixture ZWp3 that inserts respectively in each vestibule of liner, amplification buffer 4, DNA marker5 and agarose 6 form.

Use this test kit that 30 13 embryo duck in age embryos and 40 Gaoyou ducks of known sex are carried out molecule sex Classification Identification.

1, sample

Gather the chorioallantoic membrane of 30 13 embryo duck in age embryos and the blood of 40 Gaoyou ducks, use traditional phenol/chloroform method and extract DNA, and with the DNA concentration dilution to 50ng/ μ l.

2, pcr amplification reaction and result

1) the pcr amplification reaction system is prepared

2) pcr amplification reaction programming

3, the agarose electrophoresis of pcr amplification product is identified

Reaction is got 4-10 μ l reaction product at 2% agarose gel, electrophoresis in 1 * TBE electrophoresis liquid after finishing.Under the room temperature, according to electrophoresis chamber width * 6-8V, electrophoresis 20 minutes.

According to electrophoretogram, the identified gene type.The PCR product that occurs below corresponding DNA marker 500bp is that Z is with 495 bp, and the PCR product that occurs between corresponding DNA marker 250-500bp is that W is with 351 bp.Wherein, below corresponding DNA marker 500bp, occur a PCR product male ZZ type, the female ZW type that two PCR products between corresponding DNA marker 250-500bp, occur, sample individual molecular sex identification result sex confirmation known with it.

Claims

1. the quick identification reagent box of duck sex box body is characterized in that, described test kit is by box body (1), liner (2) in the box body, the Auele Specific Primer mixture ZWp(3 that inserts respectively in each vestibule of liner), amplification buffer (4), DNA marker(5) and agarose (6) composition.