A method for extracting free DNA from serum plasma
A plasma and serum technology, applied in the field of free DNA extraction, can solve the problems of poor enrichment of free DNA by the adsorption membrane, low concentration of free DNA, poor repeatability of nucleic acid, etc., and achieve a suitable extraction process for large-scale promotion and application Simple, increasing difficulty effects
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Embodiment 1
[0044] Embodiment 1, the preparation of silicon dioxide nanomaterial film:
[0045] A: Add the cage-type mesoporous silica nanomaterial into deionized water and stir evenly, then add N-carboxymethyl chitosan and stir evenly, then stir at 70°C for 4 hours, the silica nanomaterial and N-carboxylate The weight ratio of methyl chitosan is 3:2, filter, and wash 2 times with deionized water, collect powder after freeze-drying, obtain the modified silicon oxide nanomaterial;
[0046] B dissolving N-carboxymethyl chitosan in deionized medium and stirring evenly to prepare a film-forming solution of N-carboxymethyl chitosan with a mass concentration of 5%;
[0047] C, adding the modified silicon oxide nanomaterial obtained in step A to the film-forming liquid obtained in step B, the solid-liquid ratio of the modified silicon oxide nano-material to the film-forming liquid is 1g:5ml, drying, and forming a film , that is.
Embodiment 2
[0048] Embodiment 2, a kind of method extracting free DNA from serum plasma
[0049] S1 Take 200 μl of serum sample into a 1.5ml centrifuge tube, add 600 μl of Lysis Solution A, mix upside down or vortex, and place in a water bath at 65°C for 5 minutes to obtain Mixed Solution I;
[0050] The preparation method of described lysate A is: the concentration is 60mmol / LTris-Hcl, 0.3g sodium chloride, 0.8g edetate disodium, 10g resveratrol, 8g puerarin, volume concentration is 4% Alkyl glucoside, the volume concentration is that 40% isopropanol solution is added in the beaker for dissolving, transfers in the volumetric flask, the DEPC treatment water is settled to 50ml, adjusts the pH value to be 6.0, makes;
[0051] S2 Transfer the mixed solution I obtained in step S1 to an adsorption column, place it at room temperature for 5 minutes, centrifuge at a speed of 12000 rpm for 1 minute, and pour off the waste liquid in the collection tube;
[0052] S3 Put the adsorption column treat...
Embodiment 3
[0056] Embodiment 3, a kind of method for extracting free DNA from serum plasma
[0057] S1 Take 200 μl of plasma sample into a 1.5ml centrifuge tube, add 600 μl of lysate A, mix upside down or vortex, and place in a water bath at 65°C for 5 minutes to obtain mixed solution I;
[0058] The preparation method of described lysate A is: the concentration is 70mmol / LTris-Hcl, 0.5g sodium chloride, 1.2g edetate disodium, 16g resveratrol, 12g puerarin, volume concentration is 6% Alkyl glucoside, the volume concentration is that 40% isopropanol solution is added in the beaker for dissolving, transfers in the volumetric flask, the DEPC treatment water is settled to 50ml, adjusts the pH value to be 5.2, makes;
[0059] S2 Transfer the mixed solution I obtained in step S1 to an adsorption column, place it at room temperature for 5 minutes, centrifuge at a speed of 12000 rpm for 1 minute, and pour off the waste liquid in the collection tube;
[0060] S3 Put the adsorption column treated...
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