Preparation and detection method of Bacterium typhosum rapid detection reagent kit
A technology of Salmonella typhi and detection kit, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, resistance to vector-borne diseases, etc., can solve the problems of high detection sensitivity and long time required, and achieve high sensitivity , a strong specific effect
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Embodiment 1
[0033] Prepare the Loop-Mediated Isothermal Amplification Kit for Salmonella Typhi according to the following recipe:
[0034] (1) Preparation of LAMP reaction solution:
[0035] Contains 2.5 μL 10× Thermopol reaction buffer, 1.0 μL 10 mmol / L dNTP, 1.0 μL 20 μmol / L upstream internal primer (FIP), 1.0 μL 20 μmol / L downstream internal primer (BIP), 0.25 μL 20 μmol / L upstream external primer ( F3), 0.25 μL 20 μmol / L downstream outer primer (B3), 0.5 μL 100 mmol / LMgSO 4 , 12.5 μL 2mol / L betaine and 4 μL ddH 2 O.
[0036] The upstream internal primers described therein:
[0037] 5-TTACGTCACCAACGACACGGG-ACGTATTGGGCAATGACTGG-3,
[0038] Downstream inner primer: 5-CGGTAAATACCATCCCCACGGC-TAACGCAGCGAGAATGGC-3,
[0039] Upstream outer primer: 5-GTCGCGTACTTTACGCCAT-3,
[0040] Downstream outer primer: 5-ACCCTGACCATCCACCAG-3;
[0041] The mass ratio of the four deoxyribose nucleic acids in the mixture dNTP is dUTP:dATP:dGTP:dCTP=2:1:1:1.
[0042] (2) UNG enzyme: 1U / μL;
[0043](3)...
Embodiment 2
[0057] Prepare the Loop-Mediated Isothermal Amplification Kit for Salmonella Typhi according to the following recipe:
[0058] (1) Preparation of LAMP reaction solution:
[0059] Contains 2.5 μL 10× Thermopol reaction buffer, 1.0 μL 10 mmol / L dNTP, 1.0 μL 20 μmol / L upstream internal primer (FIP), 1.0 μL 20 μmol / L downstream internal primer (BIP), 0.25 μL 20 μmol / L upstream external primer ( F3), 0.25 μL 20 μmol / L downstream outer primer (B3), 0.5 μL 100 mmol / L MgSO 4 , 12.5 μL 2mol / L betaine and 4 μL ddH 2 O.
[0060] The upstream internal primer, downstream internal primer, upstream external primer, and downstream external primer are the same as above. The mass ratio of the four deoxyribose nucleic acids in the above mixture dNTP is dUTP:dATP:dGTP:dCTP=2:1:1:1.
[0061] (2) UNG enzyme: 1U / μL;
[0062] (3) Bst DNA polymerase: 8U / μL;
[0063] (4) Chromogen: 10% fluorescent dye SYBR GREEN I or DNAGreen.
[0064] Follow the procedures (1)-(3) below for testing:
[0065] (...
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