Reagent kit special for testing high pathogenicity pig replication and syndrome virus variation strain
A syndrome virus, highly pathogenic technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problem of high technical requirements, high requirements of laboratory personnel, and difficult to be applied to on-site inspection on a large scale and other problems to achieve a strong specific effect
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[0033] 1) Primers
[0034] According to the characteristic of deletion in the Nsp2 region in the highly pathogenic porcine reproductive and respiratory syndrome that occurred after 2006, a pair of specific primers were designed. The amplified fragment size is 404bp, and the amplified traditional PRRSV amplified fragment is 494bp. Synthesized by TaKaRa Biotechnology Co., Ltd.
[0035] PSX1: 5'-TGGTCCTAACGGTTCGGAAGAAAC-3'
[0036] PSX2: 5'-GTGAGCTGAGTATTTTGGGCGTGT-3'
[0037] 2) Treatment of sick materials
[0038] Cut the disease material into pieces and grind them in a sterilized grinder, dilute them 1:4 times with sterile cell culture medium, take the virus suspension at -20°C for 3 times, centrifuge at 8000rpm for 10min, and take the supernatant- Store at 80°C for later use.
[0039] 3) Extraction of viral RNA
[0040] ① Take 300L of grinding solution and add 600μL RNAout, shake and mix well and let stand at room temperature for 5min;
[0041] ② Add 200 μL of chlorofo...
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