Nano magnetic bead for nucleic acid purification
A nano-magnetic bead and magnetic bead technology, applied in the field of material science, can solve the problems of incomplete modification of surface groups, low degree of automation, and instability of magnetic beads, so as to reduce workload, realize automation, broad application prospects and social The effect of economic benefits
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Embodiment 1
[0037] Embodiment 1: Preparation of nano magnetic beads for nucleic acid purification
[0038] Step A) preparation of nano-microspheres:
[0039] Under the protection of argon, 1 ml of 0.5 M sodium hydroxide solution was added dropwise at intervals of 10 minutes to the aqueous solution containing 15 g of ferrous sulfate heptahydrate and 10 g of ferric chloride hexahydrate. When the reaction solution turns into a colloidal emulsion, add 1ml of 0.25M sodium hydroxide dropwise every 20 minutes, shake the shaker continuously at a speed of 100 rpm, and continue argon protection after the magnetic field appears. Under reaction 4~10 hours. Then, according to the volume of magnetic beads, sodium chloride was added to make the final concentration 1M. After standing at room temperature for 18-36 hours, rinse with deionized water until the pH of the solution is close to 7. Use 0.25M sodium hydroxide to prepare 30% magnetic bead suspension;
[0040] Step B) surface modification of nano ...
Embodiment 2
[0043] Embodiment 2: The real-time fluorescent quantitative PCR detection of HBVDNA
[0044] The same method as in Example 1 was used to prepare magnetic beads.
[0045] Prepare the lysate and wash solution mixed with magnetic beads according to the following component ratios:
[0046] 0.3N Sodium Hydroxide, 0.45M Potassium Chloride, 0.03% Sodium N-Lauroyl Sarcosinate, 5mM EDTA, 0.45M Tris-HCL, 1% Triton X-100, 3mMDTT and 0.001wt% Bromophenol Blue.
[0047] Add the magnetic beads that have naturally precipitated for 24 hours according to the volume ratio of magnetic beads and lysate of 1:100, and mix well.
[0048] Washing liquid: 0.3M potassium chloride, 0.001wt% stone green, adjust the pH value to 4 with acetic acid.
[0049] Use the above lysate and wash solution for clinical diagnosis and the tested HBVDNA quantitative value is 5IU / ml, 10IU / ml, 20IU / ml, 2×10 2 IU / ml, 2×10 3 IU / ml, 2×10 4 IU / ml, 2×10 5 IU / ml, 2×10 6 IU / ml, 2×10 7 IU / ml, 2×10 8 IU / ml, 2×10 9 The he...
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