Molecular biological variety identification method for Huangjiaji chicken
A molecular biology and identification method technology, applied in the field of biological species identification, can solve problems such as difficult, unscientific, accurate, and distinguishable, and achieve the effects of low cost, short identification time, and high accuracy
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Embodiment 1
[0030] The tested chicken is Huangjiaji chicken:
[0031] (1) Extract the total DNA including mitochondrial DNA from Huangjiaji chicken (1, breed code named HJJ1) and tested chicken (2 Huangjiaji chickens, breed code named SHJJ1 and SHJJ2) respectively by phenol-chloroform method;
[0032] (2) Three pairs of primers PF1 (SEQ ID NO.3), PR1 (SEQ ID NO. 4), PF2 (SEQ ID NO.5), PR2 (SEQ ID NO.6) and PF3 (SEQ ID NO.7), PR3 (SEQ ID NO.8); the sequences of the primers are respectively:
[0033] Upstream primer PF1: 5'-TACAGCCTAACGCTTCAACA-3';
[0034] Downstream primer PR1: 5'-GGTTGCGGTCGGTAAGTAGT-3'.
[0035] Upstream primer PF2: 5'-GCCCATGCTTTCGTCATAA-3';
[0036] Downstream primer PR2: 5'-TGGGATGGCGATGATTATTG-3'.
[0037] Upstream primer PF3: 5'-CGCCATCCCAACTGGTAT-3';
[0038] Downstream primer PR3: 5'-GATGAGGCGTCTTGAAAG-3'.
[0039] (3) Utilize the primers described in step (2) to carry out PCR amplification to the COI gene of Huangjiaji chicken mitochondrial DNA, the amplif...
Embodiment 2
[0055] The tested chicken is native chicken:
[0056] (1) Extract the total DNA of Huangjiaji chicken (1, breed code-named HJJ1) and tested chickens (3 native chickens, breed code-named TJ1, TJ2 and TJ3) including mitochondrial DNA by the phenol-chloroform method;
[0057] (2) Three pairs of primers PF1 (SEQ ID NO.3), PR1 (SEQ ID NO. 4), PF2 (SEQ ID NO.5), PR2 (SEQ ID NO.6) and PF3 (SEQ ID NO.7), PR3 (SEQ ID NO.8); the sequences of the primers are respectively:
[0058] Upstream primer PF1: 5'-TACAGCCTAACGCTTCAACA-3';
[0059] Downstream primer PR1: 5'-GGTTGCGGTCGGTAAGTAGT-3'.
[0060] Upstream primer PF2: 5'-GCCCATGCTTTCGTCATAA-3';
[0061] Downstream primer PR2: 5'-TGGGATGGCGATGATTATTG-3'.
[0062] Upstream primer PF3: 5'-CGCCATCCCAACTGGTAT-3';
[0063] Downstream primer PR3: 5'-GATGAGGCGTCTTGAAAG-3'.
[0064] (3) Utilize the primers described in step (2) to carry out PCR amplification to the COI gene of Huangjiaji chicken mitochondrial DNA, the amplified product is cl...
Embodiment 3
[0077] The tested chicken is black-bone chicken:
[0078] (1) Extract the total DNA of Huangjiaji chicken (1, breed code-named HJJ1) and tested chickens (3 black-bone chickens, breed code-named WJ1, WJ2 and WJ3) including mitochondrial DNA by the phenol-chloroform method;
[0079] (2) Three pairs of primers PF1 (SEQ ID NO.3), PR1 (SEQ ID NO. 4), PF2 (SEQ ID NO.5), PR2 (SEQ ID NO.6) and PF3 (SEQ ID NO.7), PR3 (SEQ ID NO.8); the sequences of the primers are respectively:
[0080] Upstream primer PF1: 5'-TACAGCCTAACGCTTCAACA-3';
[0081] Downstream primer PR1: 5'-GGTTGCGGTCGGTAAGTAGT-3'.
[0082] Upstream primer PF2: 5'-GCCCATGCTTTCGTCATAA-3';
[0083] Downstream primer PR2: 5'-TGGGATGGCGATGATTATTG-3'.
[0084] Upstream primer PF3: 5'-CGCCATCCCAACTGGTAT-3';
[0085] Downstream primer PR3: 5'-GATGAGGCGTCTTGAAAG-3'.
[0086] (3) Utilize the primers described in step (2) to carry out PCR amplification to the COI gene of Huangjiaji chicken mitochondrial DNA, the amplified product...
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