DNA extracting method for evaluating community diversity of the intestinal microorganisms of animals
A technology of intestinal microorganisms and extraction methods, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA preparation, etc., can solve the problems of difficult removal of pollutants in the purification step, low DNA recovery rate, etc., and reduce the risk of health problems. Injury, low cost, high yield effect
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Embodiment 1
[0034] (1) Add 1.5 ml of sample soaking solution A to 0.5 g of mouse feces sample, vortex for 10 minutes, centrifuge at 10,000 rpm for 5 minutes, discard the supernatant, and repeat 3 times.
[0035] (2) Add 1.5ml of sample immersion solution 2, place on ice for 5 minutes, vortex for 10 minutes, centrifuge at 10,000 rpm for 5 minutes, discard the supernatant, and repeat 3 times.
[0036] (3) Add 0.3 g of quartz sand and 1 glass bead with a diameter of 4 mm, and add 978 μl of phosphate buffer C (pH=8.0) to the sample. Vortex for 5 minutes.
[0037] (4) Add 122 μl of cell lysate D to the sample, and vortex for 5-20 minutes. Centrifuge at 13000rmp for 10min to precipitate debris.
[0038] (5) Transfer the supernatant to a clean centrifuge tube, add 250 μl of protein removal solution E to the tube, gently invert and mix 10 times by hand, leave at room temperature for 5 minutes, and centrifuge at 13,000 rpm for 5 minutes.
[0039] (6) Transfer the supernatant to a clean centrifu...
Embodiment 2
[0048] (1) Add 1.5ml sample soaking solution A to 0.5g rat feces sample, vortex for 10 minutes, 10000 rpm, centrifuge for 5 minutes, discard the supernatant, repeat 3 times.
[0049] (2) Add 1.5ml sample immersion solution 2, place on ice for 5 minutes, vortex for 10 minutes, 10000 rpm, centrifuge for 5 minutes, discard the supernatant, repeat 3 times.
[0050] (3) Add 0.3 g of quartz sand and 1 glass bead with a diameter of 4 mm, and add 978 μl of phosphate buffer C (pH=8.0) to the sample. Vortex for 5 minutes.
[0051] (4) Add 122 μl of cell lysate D to the sample, and vortex for 5-20 minutes. Centrifuge at 13000rmp for 10min to precipitate debris.
[0052] (5) Transfer the supernatant to a clean centrifuge tube, add 250 μl of protein removal solution E to the tube, gently invert and mix 10 times by hand, leave at room temperature for 5 minutes, and centrifuge at 13,000 rpm for 5 minutes.
[0053] (6) Transfer the supernatant to a clean centrifuge tube, add 3 times the vo...
Embodiment 3
[0062] (1) Add 1.5ml sample soaking solution A to 0.5g rabbit feces sample, vortex for 10 minutes, 10000 rpm, centrifuge for 5 minutes, discard the supernatant, repeat 3 times.
[0063] (2) Add 1.5ml sample immersion solution 2, place on ice for 5 minutes, vortex for 10 minutes, 10000 rpm, centrifuge for 5 minutes, discard the supernatant, repeat 3 times.
[0064] (3) Add 0.3 g of quartz sand and 1 glass bead with a diameter of 4 mm, and add 978 μl of phosphate buffer C (pH=8.0) to the sample. Vortex for 5 minutes.
[0065] (4) Add 122 μl of cell lysate D to the sample, and vortex for 5-20 minutes. Centrifuge at 13000rmp for 10min to precipitate debris.
[0066] (5) Transfer the supernatant to a clean centrifuge tube, add 250 μl of protein removal solution E to the tube, gently invert and mix 10 times by hand, leave at room temperature for 5 minutes, and centrifuge at 13,000 rpm for 5 minutes.
[0067] (6) Transfer the supernatant to a clean centrifuge tube, add 3 times the...
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