Primer pair and kit used for carrying out early stage gender authentication of chicken, and application of primer pair and kit
A kit and a technology of specific primer pairs, applied in the field of primer pairs for early sex identification of chickens, can solve the problems of easy misjudgment, cumbersome experimental process, high toxicity, etc.
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Embodiment 1
[0030] Embodiment 1, establishment of method
[0031] Samples for testing: 10 adult camellia chickens of known sex, 5 of which are numbered 1-5 are roosters, and 5 of which are numbered 6-10 are hens.
[0032] 1. Take the blood of the test sample and extract the genomic DNA.
[0033] 2. Using the genomic DNA obtained in step 1 as template DNA, perform PCR amplification.
[0034] The primer pairs used in PCR amplification are as follows:
[0035] Forward primer (SEQ ID NO.1): 5'-TGTATTTTGGTTCTACAGGC-3';
[0036] Reverse primer (SEQ ID NO. 2): 5'-ATCATCATAACCATAAATCT-3'.
[0037] PCR amplification reaction system (50 μL): 2×PCR Mix (Nanjing Boerdi Biological Co., Ltd.) 25 μL, 10 μmol / L forward primer 1 μL, 10 μmol / L reverse primer 1 μL, 50-100 μg / ml template DNA 2 μL, Ultrapure water 21 μL.
[0038] The reaction program of PCR amplification: 95°C for 5min; 95°C for 30s, 60°C for 30s, 72°C for 30s, 35 cycles; 72°C for 10min.
[0039] 3. After step 2 is completed, the PCR am...
Embodiment 2
[0042] Embodiment 2, the application of method
[0043] Samples for testing: 17 Tibetan chicken chicks of unknown sex.
[0044] Method is with embodiment 1.
[0045] Electropherogram see figure 2 . figure 2 In , lanes 1 to 17 sequentially represent 17 samples for testing. Among them, some of the test samples showed a single characteristic band of 538bp (swimming lanes 1, 3, 6, 7, 8, 10, 11, 15, 17), which were judged to be roosters, and some of the test samples showed two characteristic bands of 960bp and 538bp (Swimming lanes 2, 4, 5, 9, 12, 13, 14, 16), judged to be a hen.
[0046] The test specimens were dissected and sexual organs were observed, and the above identification results were completely correct.
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