High-precision nucleic acid quantitative detection kit for hepatitis C virus (HCV)
A hepatitis C virus, detection kit technology, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of poor accuracy, low sensitivity, large sample size, etc., to eliminate interference and achieve good repeatability , a strong specific effect
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[0025] Examples of fluorescence quantitative PCR detection of HCV RNA
[0026] (1) Reagent preparation
[0027] a. Preparation of RT-PCR reaction solution
[0028] The RT-PCR buffer composition is 50mmol / L Tris-HCl (PH8.3), 10mmol / L (NH4) 2 SO 4 , 75mmol / LKCl, 2.5mmol / L MgSO 4 , 0.15mmol / L dNTPs. The upstream and downstream primers used to amplify the target gene were used at a concentration of 20 pmol, the target gene detection probe used at a concentration of 8 pmol, the internal standard upstream primer and downstream primer used at a concentration of 8 pmol, and the internal standard probe used at a concentration of 8 pmol.
[0029] b. Take the corresponding amount of lysis solution binding solution and internal standard according to the proportion (lysis binding solution 400μl / person + internal standard 0.1μl / person portion), mix thoroughly to form the lysis binding solution-mix, and then centrifuge for use.
[0030] c. According to the amount of test sample, negative control, pos...
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