Molecular marker influencing chicken shin skin color and application of molecular marker
A technology of molecular marking and skin color, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of inability to identify skin color, reduce breeding time, promote sales, and increase sales Effect
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Embodiment 1
[0036] Example 1 Primer Design
[0037] 1. Experimental method
[0038] Find the sequence of the 3' non-coding region of the ALDH1A3 gene from https: / / www.ncbi.nlm.nih.gov / , and design a primer pair. The primer pair information is shown in Table 1 (primer sequence 5'→3'). The sequence of the primer pair was sent to Guangzhou Sangon Biotech Co., Ltd. for synthesis, and the primer pair was used to perform PCR on tissue DNA to test the specificity of the primers.
[0039] Table 1 Primer information for SNP screening of ALDH1A3
[0040]
[0041] 2. Experimental results
[0042] The results are attached figure 1 shown. Use this primer pair to carry out PCR on blood DNA, as attached figure 1 As shown, the primers are specific
Embodiment 2
[0043] Example 2 DNA pool sequencing
[0044] 1. Experimental method
[0045] The sample chickens were selected from 600 120-day-old Nanhai Jute Chicken No. 1 pure line of Jiangfeng Chicken Farm. All chickens were hatched in the same batch, raised in a flat way, and fed with corn-soybean meal that meets international formula standards feed. Use a 2ml syringe to collect blood from the subwing vein, collect the blood and store it in a centrifuge tube containing 20% EDTA for blood DNA extraction.
[0046] Genomic DNA of all individuals based on Plant Mini Kit (Qiagen, Hilden, CA; Cat#69104) was used in the kit instructions to extract blood DNA, measure the quality and concentration, dilute to 50 ng / μL, and store at 4°C for later use.
[0047] 30 DNA samples were randomly selected from the total extracted DNA samples to construct a mixed pool, and every three samples (about 0.33 μL of each sample) were mixed into a mixed sample, a total of 10 samples. The mixed pool samples ...
Embodiment 3
[0050] Example 3 Determination of genotype determination of ALDH1A3 mutation site g.703T>C
[0051] 1. Experimental method
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