Reagent box for detecting prawn infectious subcutaneous and hemopoietic tissue necrosis virus and method thereof
A technology of hematopoietic tissue and necrotic virus, which is applied in the direction of biochemical equipment and methods, and microbial measurement/inspection, can solve the problems of low sensitivity, complicated operation, high cost, etc., and achieve high sensitivity and strong specificity
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[0051] The following examples are further illustrations of the present invention and should not be regarded as limiting the present invention.
[0052] Prepare the isothermal branch amplification detection kit for shrimp infectious subcutaneous and hematopoietic necrosis virus according to the following formula: tissue lysate A: 4MGTC, 25mM sodium citrate, 0.5% sodium lauryl sarcosine and 0.1M β-mercaptoethanol DNA adsorption solution B: 50% Golden Beads suspension
[0053] Hybridization solution C: 2M GTC, 0.5% BSA, 80mM EDTA, 400mM Tris-HCl pH7.5, 0.5% sodium lauryl sarcosine, 2.5nM IHHNV-C-probe and 25nM IHHNV-capture probe
[0054] Magnetic beads D: 4mg / ml SAV immunomagnetic beads
[0055] Probe capture solution E: 10mM Tris-HCl pH7.5, 1mM EDTA and 2M NaCl
[0056] Wash solution F: 10 mM Tris-HCl pH 8.0 and 1 mM EDTA
[0057] Ligation solution G: 20ul 1×ligase buffer and 0.25ul Taq DNA ligase (40U / ul)
[0058] RAM reaction solution H: 6ul 2.5mM dNTP, 5ul 10×Thermopol r...
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