Method and kit for rapidly extracting nucleic acids from trace clinical samples
A sample-in-kit technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA preparation, etc., can solve the problems of inability to completely remove PCR inhibitors, high DNA purity, complicated steps, etc., and achieve operation time. Short, easy to operate, low cost effect
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Embodiment 1
[0066] Sputum processing and TB DNA extraction
[0067] 1. Sputum liquefaction
[0068] Take the sputum of the tubercle bacillus (TB) subject to be tested, transfer it to a centrifuge tube, add 1 times the volume of sputum liquefaction solution, shake and mix well, heat at 95°C-100°C for 10 minutes, and cool to room temperature;
[0069] 2. Lysis
[0070] Take 1ml of liquefied sputum, add the same amount of lysate; mix well and let it stand at room temperature for 5 minutes; use the micro-clinical sample nucleic acid rapid extraction device to suck up the above mixed solution, and push out the filtrate in the suction device through a pointed pipette and discard Lose;
[0071] 3. Cleaning
[0072] Use the micro-clinical sample nucleic acid rapid extraction device to suck in the centrifuge tube containing the cleaning solution, and then push out the filtrate in the suction device through a pointed suction pipe and discard it;
[0073] 4. DNA Elution
[0074] Continue to use...
Embodiment 2
[0086] Treatment of sexually transmitted disease samples (cotton swabs) and extraction of pathogen DNA
[0087] 1. Elution
[0088] Take a cotton swab sample from a Chlamydia trachomatis (CT) patient to be tested, add 1ml of normal saline to the sample collection tube, shake and mix well to ensure that the secretions on the cotton swab are fully eluted;
[0089] 2. Lysis
[0090] Add an equal amount of lysate to the above eluent; mix well and let stand at room temperature for 5 minutes; use a micro-clinical sample nucleic acid rapid extraction device to suck up the above mixed solution, and push out the filtrate in the suction device through a pointed pipette and discard;
[0091] 3. Cleaning
[0092] Use the micro-clinical sample nucleic acid rapid extraction device to suck in the centrifuge tube containing the cleaning solution, and then push out the filtrate in the suction device through a pointed suction pipe and discard it;
[0093] 4. DNA Elution
[0094] Continue to...
Embodiment 3
[0108]Serum sample processing and extraction of hepatitis C virus (HCV) RNA
[0109] 1. Lysis
[0110] Take 200ul of HCV serum sample (thaw the frozen serum at room temperature before use, shake and mix for 10 seconds), add an equal amount of lysate, mix well, and let stand at room temperature for 5 minutes; use the micro-clinical sample nucleic acid rapid extraction device to absorb the above mixed Liquid, the filtrate in the suction device is pushed out through the pointed suction pipe and discarded;
[0111] 2. Cleaning
[0112] Use the micro-clinical sample nucleic acid rapid extraction device to suck in the centrifuge tube containing the cleaning solution, and then push out the filtrate in the suction device through a pointed suction pipe and discard it;
[0113] 3. RNA Elution
[0114] Continue to use the micro-clinical sample nucleic acid rapid extraction device to suck 100ul of the eluent, and push the liquid in the suction device into a new centrifuge tube through ...
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