Alternaria solani Sorauer detection kit and detection method thereof
A detection kit and technology for early blight bacteria, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the long period of sporulation induced by early blight bacteria in vitro, difficult to adapt to species identification, leaf and tissue symptoms problems such as slow development
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[0013] (1) Kit components include: primers AsF and AsR (10 μM each), Mix [Taq enzyme (0.1-0.5 U / μL), dATP (0.05-0.10 mM), dTTP (0.05-0.10 mM), dGTP (0.05 -0.10 mM), dCTP (0.05-0.10 mM), MgCl 2 (0.5-1.0 mM), KCl (100-150 mM), Tris-HCl (20-25 mM, pH 9.0-9.3), 1% Triton X-100 and stabilizers], ddH 2 O;
[0014] (2) Extraction of DNA: the hyphae of Phytophthora infestans was extracted by CTAB method, and the leaves in the field were extracted by improved glass bead shaking method;
[0015] (3) Polymerase chain reaction:
[0016] A. Polymerase chain reaction takes 25ul as a reference, and the dosages of various items are:
[0017] Template DNA 1.0 μL
[0018] Primer-AsF 1.0 μL
[0019] Primer-AsR 1.0 μL
[0020] Mix 12.5μL
[0021] wxya 2 O 9.5 μL
[0022] B. PCR reaction conditions: The reaction is carried out on a PCR instrument, and the reaction conditions are:
[0023] Initial denaturation stage: 95°C, 1min;
[0024] Denaturation stage: 95°C, 25-35s;
[0025...
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