Polymerase chain reaction (PCR) fluorescence detection kit and detection method for candida albicans
A Candida albicans and fluorescence detection technology, applied in the field of microbial detection, can solve the problems of poor repeatability, many influencing factors, low resolution, etc., and achieve the effects of short time consumption, simple operation, high sensitivity and specificity
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Embodiment 1
[0033] The composition of embodiment 1 kit of the present invention
[0034] Main raw material sources and preparation methods:
[0035] Tris: analytically pure, a product from a qualified supplier, content 99.7%, infrared qualified, pH (5% water) 10.3-10.9, moisture 0.3%, melting point 167-171°C, qualified absorption system, qualified for the highest content of impurities .
[0036] MgCl 2 : Analytical pure, the product of a qualified supplier, the content is not less than 99%, the aqueous solution reaction is qualified, and the highest content of impurities is qualified. (MgCl 2 Very easy to absorb moisture, put it in a desiccator after using the new bottle).
[0037]EDTA: Analytical pure, the product of a qualified supplier, it is a white crystalline powder, soluble in water, the solution is acidic, hardly soluble in alcohol, the content is not less than 99.5%, the aqueous solution reaction is qualified, and the complexing force test is qualified. The highest content o...
Embodiment 2
[0080] Embodiment 2 The use of kit of the present invention
[0081] 1. Reagent preparation:
[0082] 1. Take out 10× concentrated cleaning solution A and 10× concentrated cleaning solution B, dilute them with sterile pure water at a ratio of 1:9 (volume ratio), and put them in a 4°C refrigerator for later use.
[0083] 2. Centrifuge Taq DNA Polymerase and Uracil N-Glycosylase (UNG) briefly, and put them in a -20°C refrigerator for later use.
[0084] 3. After determining the number of reaction tubes n (number of samples + negative and positive controls), take out the CA-PCR reaction solution, and mix n×44.3μl CA-PCR reaction solution, n×0.5μl Taq DNA Polymerase and n×0.2μl Add Uracil N-Glycosylase (UNG) into a centrifuge tube and oscillate to mix well. After centrifuging briefly, dispense 45 μl into each PCR reaction tube, cover the tube cap and transfer to the sample loading area, and store in a 4°C refrigerator in the dark. spare.
[0085] 4. Transfer the extracted solid...
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