A Molecular Marker for Identifying Cherry Valley Duck and Its Application
A technology of Cherry Valley Duck and varieties, applied in the field of molecular biology, can solve problems such as difficulty in rights protection, shortage, and difficulty in identification, and achieve the effects of high accuracy, fast speed, and easy operation
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Embodiment 1
[0045] Example 1 Identification of Cherry Valley Duck SNP Molecular Marker Development
[0046] A total of 7 duck species including Cherry Valley Duck, Peking Duck, Maple Leaf Duck, Jinding Duck, Shaoxing Duck, Shanma Duck and Gaoyou Duck were selected as the source of experimental materials, regardless of sex, and 8 individuals for each duck species.
[0047] 1. Whole genome resequencing:
[0048] Blood was collected from the subwing vein of the test ducks, and DNA was extracted by the phenol-chloroform method. Each DNA sample was sequenced by Illumina Hiseq 2500, and the average sequencing depth of each individual was 6.45X. A total of 376.06Gb reads with a paired-end length of 100bp were generated by sequencing.
[0049]2. Reads assembly and mutation search:
[0050] Use NGS QC Toolkit (default parameters) to filter low-depth reads to avoid human errors during library construction and sequencing, and compare the filtered reads to the duck reference genome Anasplatyrhyncho...
Embodiment 2
[0060] Example 2 is used to identify the application of the SNP molecular marker of Cherry Valley Duck (1)
[0061] 1. Extract the genomic DNA of the ducks to be tested. After the DNA of 8 ducks is extracted from each breed of duck and mixed, the mixed sample is used as the genomic DNA of each breed of ducks to be tested.
[0062] 2. Amplify the nucleotide fragment containing the SNP site
[0063] For the SNP site (KB743619.1:351724) screened in Example 1, according to the first pair of primers in Table 2, using genomic DNA as a template, PCR is carried out, and the amplification system used in the PCR amplification reaction, when the system 25ul The preparation is as follows: each 0.5ul of upstream and downstream primers, concentration 10pmol / L; ddH 2 O 10.5ul; 2×Taq PCR Mix 12.5ul; DNA template 1ul, concentration 100ng / ul;
[0064] Amplification program: pre-denaturation at 94°C for 5 minutes, 35 cycles at 94°C for 30s, 60°C for 40s, 72°C for 30s, and finally extension at ...
Embodiment 3
[0069] Example 3 is used to identify the application of the SNP molecular marker of Cherry Valley Duck (2)
[0070] 1. Extract the genomic DNA of the ducks to be tested. After the DNA of 8 ducks is extracted from each breed of duck and mixed, the mixed sample is used as the genomic DNA of each breed of ducks to be tested.
[0071] 2. Amplify the nucleotide fragment containing the SNP site
[0072] For the SNP site (KB743619.1:351767) screened in Example 1, according to the second pair of primers in Table 2, using genomic DNA as a template, PCR is carried out, and the amplification system used in the PCR amplification reaction, when the system 25ul The preparation is as follows: each 0.5ul of upstream and downstream primers, concentration 10pmol / L; ddH 2 O 10.5ul; 2×Taq PCR Mix 12.5ul; DNA template 1ul, concentration 100ng / ul;
[0073] Amplification program: pre-denaturation at 94°C for 5 minutes, 35 cycles at 94°C for 30s, 60°C for 40s, 72°C for 30s, and finally extension at...
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