Nucleic acid releaser and method for quickly extracting nucleic acids from dried blood spots
The technology of a nucleic acid releasing agent and an extraction method is applied in the field of nucleic acid releasing agent and fast extraction of nucleic acid from dried blood spots, which can solve the problems such as inability to effectively remove impurities such as hemoglobin, and achieve the effects of low cost, easy identification and convenient operation.
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Embodiment 1
[0073] Preparation of nucleic acid release agent: Tris-HCl, EDTA, MgCl with a pH value of 7.4 2 , TrixonX-100, KCL and proteinase K were added into sterile water and mixed evenly, among which Tris-HCl, EDTA, MgCl 2 , TrixonX-100, KCL, and Proteinase K are at concentrations of 13mM / L, 2mM / L, 14.5mM / L, 2mM / L, 10mM / L, and 0.065mg / mL, and are stored at -20°C;
[0074] Preparation of blood spot sample discs: Separate a 5×5mm sample disc from the dried blood spot collection card, put it into a centrifuge tube, and use a puncher to punch a 5×5mm blood spot (or a round A blood spot cut into a sample of approximately 5×5 mm).
[0075] The nucleic acid release solution was transferred to a filter column, and centrifuged at a centrifugal force of 5000×g for 1 min to obtain a filtrate.
[0076] Transfer the filtrate to another clean, enzyme-free centrifuge tube and store it at -20°C to obtain the finally extracted and purified dried blood spot nucleic acid release agent. The parameters ...
Embodiment 2
[0091] 1L solution of nucleic acid release agent, in which Tris-HCl, EDTA, MgCl 2 , TrixonX-100, KCL and proteinase K concentrations were 10mM / L, 2.5mM / L, 10mM / L, 5mM / L, 14.005mM / L, 0.06mg / mL.
Embodiment 3
[0093] 1L solution of nucleic acid release agent, in which Tris-HCl, EDTA, MgCl 2 , TrixonX-100, KCL and proteinase K concentrations were 20mM / L, 1mM / L, 12mM / L, 1mM / L, 7.485mM / L, 0.08mg / mL.
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