Hepatitis B virus enrichment fluorescent PCR (polymerase chain reaction) detection method
A hepatitis B virus and detection method technology, applied in the field of molecular biology, can solve the problems of long PCR detection time, high nucleic acid purity requirements, and high automation cost, and achieve the effects of improving detection sensitivity, short detection time, and easy operation.
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[0039] A kind of hepatitis B virus enrichment fluorescent PCR detection method, the process flow chart of this method is as follows figure 1 As shown, the specific steps are:
[0040] 1. Preparation of immunomagnetic beads
[0041] (1) activation
[0042] Take 10 mg (25 mg / mL) fully shaken carboxy magnetic beads and place them in a 2 mL centrifuge tube, add 1 ml of 15 mMMES buffer (pH6.0) to wash the magnetic beads for 3 times; after separating with a magnetic separator, add 100 μL EDC solution (prepared with 15mM MES buffer solution (pH6.0)) with a concentration of 10mg / ml was mixed by a vortex mixer, fixed on the mixer, and activated at room temperature for 30min; the magnetic beads were separated by a magnetic separator, and discarded. Remove the supernatant.
[0043] (2) Coupling
[0044] Add 400 μg hepatitis B virus surface antibody, mix and resuspend the magnetic beads, place the centrifuge tube on a horizontal shaker, and mix at room temperature for 3 hours (2-4 hou...
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