Method for purifying DNA by using gold magnetism particles
A gold magnetic particle and magnetic separation technology, which is applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of low DNA purification efficiency, long operation time, and poor purity, and achieve large fixed capacity, long drying time, and high purification rate Effect
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Embodiment 1
[0029] Embodiment 1 is the specific process of rapid purification of genomic DNA from whole blood:
[0030] 1] Pretreatment of gold magnetic particles:
[0031] 1.1] Fully mix the gold magnetic particles to ensure that the magnetic particles are completely suspended in the solution;
[0032] 1.2] Take 100 μl of gold magnetic particles in a 1.5ml centrifuge tube, place the centrifuge tube on a magnetic separator for magnetic separation until the supernatant is clear; the separation can be completed within 1 min, and the supernatant is removed;
[0033] 1.3] Separate the centrifuge tube from the magnetic separator. Add 200 μl of pretreatment buffer to the tube, mix with a pipette, then put it back on the magnetic separator, and remove the supernatant;
[0034] 1.4] Repeat steps 1.3] to 1.4] twice. Then add 200 μl coupling buffer to the centrifuge tube and mix thoroughly.
[0035] 2] DNA purification steps:
[0036] 2.1] Take 200 μl whole blood and add it to a 1.5ml centrifu...
Embodiment 2
[0045] Embodiment 2 is the specific process of purifying plasmid DNA from bacteria:
[0046] 1] Pretreatment of gold magnetic particles:
[0047] 1.1] Fully mix the gold magnetic particles to ensure that the magnetic particles are completely suspended in the solution;
[0048] 1.2] Take 500 μl of gold magnetic particles in a 1.5ml centrifuge tube, place the centrifuge tube on a magnetic separator for magnetic separation until the supernatant is clear; the separation can be completed within 1 min, and the supernatant is removed;
[0049] 1.3] Separate the centrifuge tube from the magnetic separator. Add 200 μl of pretreatment buffer to the tube, mix with a pipette, then put it back on the magnetic separator, and remove the supernatant;
[0050] 1.4] Repeat steps 1.3] to 1.4] twice; then add 200 μl coupling buffer to the centrifuge tube and mix well;
[0051] 2] DNA purification steps:
[0052] 2.1] Take 1ml of the overnight culture solution, centrifuge at the highest speed ...
Embodiment 3
[0060] Embodiment 3 is the specific process of purifying genomic DNA from tissue:
[0061] 1.1] Take 10mg~100mg tissue high-speed homogenate and add it to a 1.5ml centrifuge tube;
[0062] 1.2] Add 30 μl ~ 300 μl lysis buffer I and 60 μl ~ 600 μl lysis buffer II to the tube, then add 1 μl ~ 10 μl RNase A solution, blow and mix with a pipette; stand at room temperature for 5 minutes;
[0063] 1.3] Add 45 μl ~ 450 μl lysis buffer III to the tube, and mix with a pipette; ice bath for 10 minutes;
[0064] 1.4] Add the pretreated gold magnetic particles into the tube, gently blow and mix with a pipette; let it stand at room temperature for 3 minutes; place the centrifuge tube on a magnetic separator for magnetic separation for 5 minutes, and remove the supernatant;
[0065] 1.5] Remove the centrifuge tube from the magnetic separator; add 200 μl ~ 1000 μl of washing buffer to the tube and mix with a pipette. Place the centrifuge tube back on the magnetic separator for magnetic sep...
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