Molecular marker influencing oblique length of chicken body and application thereof
A molecular marker and body oblique length technology, which is applied to the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc. Efficiency, the effect of reducing breeding costs
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experiment example 1
[0031] Experimental example 1 Experimental chicken group feeding and body slant length measurement
[0032] (1) Experimental animals
[0033] The test chicken group used in the present invention is the F2 generation resource group obtained by crossing Daweishan miniature chicken and recessive White Rock broiler chicken.
[0034] In the F0 generation, breeders were selected from Daweishan miniature chickens (from the practice chicken farm of Yunnan Agricultural University) and recessive white lock broilers (from Kunming Yunling Guangda Breeder Feed Co., Ltd.), and the selected male and female chickens had this breed Features, high egg production, medium weight, pure blood. Daweishan miniature chicken is a small chicken breed that has not been selected for breeding. It has the advantages of small size (average weight 0.7-1.2kg), less feed consumption, and low basal metabolism. It is a new breed of high-quality and grain-saving chickens (line ) of excellent material. Recessive...
experiment example 2
[0040] Experimental Example 2: Acquisition of Molecular Markers Affecting Chicken Body Slant Length
[0041] (1) Blood sample collection
[0042] Vacuum blood collection tubes containing EDTA-dipotassium were used to collect blood from chicken wing veins.
[0043] (2) Extraction and identification of genomic DNA
[0044] Gentra Puregene Blood Kit (Qiagen) column centrifugation kit was used to extract blood genomic DNA. After the DNA was completely dissolved, its concentration was detected with a NannoDrop nucleic acid analyzer, and the integrity of the DNA band was detected under 1% agarose gel electrophoresis conditions. sex.
[0045] (3) DNA library construction and sequencing
[0046]The construction of the library was completed using the NEBNext DNA library kit (NewEngland Biolabs). The specific operation steps were as follows: firstly, the genomic DNA was randomly broken into small fragments, and then Illumina paired-end adapters were connected to the fragments with a ...
experiment example 3
[0063] Experimental example 3 A kind of SNP molecular marker genotyping detection method that affects chicken body slant length
[0064] (1) Experimental animals
[0065] The test flock group used in the present invention is 300 (150 male and female) of 150-day-old Daweishan miniature chicken breeders of Yunnan Agricultural University, which is the core group of breeder chickens. Measure body oblique length.
[0066] (2) Target DNA sequence amplification and sequencing
[0067] 1) Primer design
[0068] The DNA sequence of chicken chromosome 5 was downloaded from the NCBI website, and primers were designed using the primer design software primer premier 6.0.
[0069] The DNA sequences of the designed primers are as follows:
[0070] Upstream primer primer-F: 5'-GAAGGTCCTCAGGGTACGTC-3';
[0071] Downstream primer primer-R: 5'-AGCCTCACGCAACAGATGAA-3'.
[0072] 2) PCR amplification
[0073] Add 1.0μL DNA template, 10.5uL double distilled water, 2×TSINGKE to the 10uL reacti...
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