Application of specific molecular marker in identifying Xueshan chicken, and kit and method for identifying Xueshan chicken
A snow mountain grass chicken and molecular marker technology, applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., can solve the problems of incorrect classification and identification, failure to identify hidden taxa, etc., and achieve duplication Highly specific, highly specific, and easy-to-operate effects
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] The present invention provides the application of a specific molecular marker in identification of snow mountain chickens, the molecular marker includes the nucleotide sequence shown in SEQ ID NO:1. The molecular marker is derived from the 3'UTR region of the BRD2 gene of Chromosome 16 of Snow Mountain Hen. The nucleotide sequence of the upstream primer for amplifying the molecular marker is shown in SEQ ID NO: 2; the nucleotide sequence of the downstream primer for amplifying the molecular marker is shown in SEQ ID NO: 3.
[0027] Table 1 shows the gene fragments of the 3'UTR region of the BRD2 gene on chromosome 16 between the Xueshan grass chicken and other chicken breeds. Due to the obvious differences between the Xueshan grass chicken and other chicken breeds in the 3'UTR region of the BRD2 gene on chromosome 16, The nucleotide sequence shown in SEQ ID NO: 1 is a unique nucleotide sequence of the Snow Mountain grass chicken, so the nucleotide sequence of the Snow M...
Embodiment 2
[0031] A kit for identifying snow mountain grass chickens, said kit comprising the following components:
[0032] DNA extraction reagent: total system 1000 μL, 0.02 mg of CTAB, 100 μL of Tris-HCL (pH8.0) with a concentration of 100 mmol / L, 200 μL of EDTA with a concentration of 1 mol / L (pH8.0), 150 μL of a concentration of 2.5 mmol / L NaCl, 0.03mg of PVP-40, the balance is ddH 2 O;
[0033] Identification system: The total system is 20 μL: 1 μL DNA of the chicken species to be identified with a concentration of 100 ng / μL, 2 μL 10×PCRBuffer, 1.5 μL dNTP with a concentration of 10 mmol / L, 1 μL each of upstream and downstream primers with a concentration of 10 pmol / μL, 0.2 μL of Taq enzyme at a concentration of 5U / μL, ddH 2 O 13.3 μL;
[0034] Positive control: The total system is 20 μL: 1 μL of DNA from snow mountain grass chicken with a concentration of 100 ng / μL, 2 μL of 10×PCRBuffer, 1.5 μL of dNTP with a concentration of 10 mmol / L, 1 μL of upstream and downstream primers w...
Embodiment 3
[0039] In order to illustrate the method for identifying snow mountain grass chickens of the present invention, this embodiment selects snow mountain grass chickens, Ross chickens, game cocks, silky chickens, Luyuan chickens, Anka chickens, You chickens, Wenchang chickens, Taihu chickens, and Liyang chickens, ten different chickens The chicken breeds were verified as the chicken breeds to be identified, and the above chicken breeds were all from the Genetic Resources Laboratory of the College of Animal Science and Technology, Yangzhou University.
[0040] 1) Collect 10 samples for each chicken breed, collect 0.4 mL of blood from the wing vein, add 0.5 mol / L Na 2 EDTA anticoagulant 2 μL, after adding 4 mL of lysate, extract DNA by phenol-chloroform extraction method; the specific steps of phenol-chloroform extraction method are as follows: after adding lysate, then add proteinase K with a final concentration of 20 mg / mL and RNaseA (ribonuclease A) with a final concentration of ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More