Nano magnetic bead based biological sample nucleic acid extracting and purifying method
A technology of nano-magnetic beads and biological samples, which is applied in the direction of DNA preparation and recombinant DNA technology. It can solve the problems of easy loss of small fragments, long time-consuming nucleic acid, and difficulty in automation, so as to shorten the extraction and purification time and improve the nucleic acid extraction process. , easily automated effects
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Embodiment 1
[0066] Embodiment 1, extracting nucleic acid from a small amount of plant cells, it is mainly implemented according to the following steps:
[0067] Step 1, plant sample preparation: wash the floating soil on the surface with sterilized water, drain quickly, weigh 5-10mg young leaves, 5-10mg petals, 5-10mg seeds, 20-40mg stems or 50-100mg young roots , put it into a 1.5ml clean centrifuge tube for later use, and then carry out the following process:
[0068] (1) Add about 500 μl of plant cell lysate into 1.5ml centrifuge tube 1, assist in breaking plant cells by physical, chemical or enzymatic methods, achieve sufficient crushing and grinding, and release nucleic acid from plant sample cells, thereby obtaining lysed cells solution;
[0069] (2) Heat the 1.5ml centrifuge tube 1 in step 1 (1) in a water bath at 70°C for 10 minutes, and perform intermittent mixing;
[0070] (3) Add 100 μl of polysaccharide-protein removal solution and 100 μl of absolute ethanol to the 1.5ml cen...
Embodiment 2
[0091] Embodiment 2, extracting nucleic acid from a small amount of animal cells, it is mainly implemented according to the following steps:
[0092] Step 1, animal sample preparation: Weigh 5mg of liver, 5-10mg of muscle, or 5-10mg of heart, put them into a 1.5ml clean centrifuge tube for later use, and then carry out the following process:
[0093] (1) Add about 600 μl of animal cell lysate into 1.5ml centrifuge tube 1, assist in breaking animal cells by physical, chemical or enzymatic methods, achieve sufficient crushing and grinding, and release nucleic acid from animal sample cells, thereby obtaining lysed cells solution;
[0094] (2) Heat the 1.5ml centrifuge tube 1 in step 1 (1) in a water bath at 50°C for 10 minutes, and perform intermittent mixing;
[0095] (3) Add 120 μl of polysaccharide-protein removal solution and 120 μl of absolute ethanol to the 1.5ml centrifuge tube 1 in step 1 (2), and mix well;
[0096] (4) Place the 1.5ml centrifuge tube in step 1(3) at ro...
Embodiment 3
[0116] Embodiment three, by extracting nucleic acid from trace bacterial cells, it is mainly implemented according to the following steps:
[0117] Step 1, Bacterial sample preparation: Cultivate the bacteria overnight, centrifuge the bacterial solution at a speed of 7000-8000r / min for 3-5 minutes, discard the supernatant, take 5-10mg of bacterial cells into a clean centrifuge tube, and then proceed to the next step Described process:
[0118] (1) Add about 600 μl of bacterial cell lysate to the 1.5ml centrifuge tube 1, and assist in breaking the bacterial cells by physical, chemical or enzymatic methods to achieve sufficient crushing and grinding, so that the bacterial sample cells release nucleic acids, thereby obtaining lysed cells solution;
[0119] (2) Heat the 1.5ml centrifuge tube 1 in step 1 (1) in a water bath at 70°C for 10 minutes, and perform intermittent mixing;
[0120] (3) Add 100 μl of polysaccharide-protein removal solution and 100 μl of absolute ethanol to ...
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