Kit for extracting nucleic acid from sputum and extraction method
A technology for extracting nucleic acid and an extraction method, which is applied in the field of biology, can solve the problems of difficult separation of cell components, abnormal cell shape variation, and difficulty in extracting high-purity DNA, and achieve simple and convenient operation steps, stable quality, and high nucleic acid purity. Effect
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Embodiment 1
[0035] Take 1 portion of sputum from a patient with tonsillitis, add 2 times the amount of digestive juice KB, shake and mix evenly. Add 50 μL proteinase K solution, 55 ° C water bath for 2 h. Add the same amount of binding solution BS as the sputum, shake and mix evenly, and then centrifuge the whole solution to pass through the nucleic acid adsorption column (pass through the column in batches, 700 μL each time). Add 500 μL SWB1 to the adsorption column, centrifuge at 12000 rpm for 1 min, then add 500 μL SWB2, centrifuge at 12000 rpm for 1 min, and finally elute with 50 μL eluent to obtain a DNA solution.
[0036] Take 5 μL of DNA solution and mix it evenly with 6×DNA Loading Buffer, add it to the sample hole of 0.7% agarose gel, electrophoresis at 5V / cm for 30 minutes, and observe the electrophoresis results under the gel imager, see figure 2 . The bands extracted from sputum can be seen in the electrophoresis diagram, and the molecular weight is greater than 23Kb.
Embodiment 2
[0038] Take the sputum of a patient diagnosed with lung cancer (the sputum sample coughed up from the lungs by the patient), add 3 times the amount of digestive juice KB, and shake and mix well. Add 100 μL of proteinase K and bathe in water at 55°C for 3h. Add the same amount of binding solution BS as the sputum, shake and mix evenly, and then centrifuge the whole solution to pass through the nucleic acid adsorption column (pass through the column in batches, 700 μL each time). Add 500 μL SWB1 to the adsorption column, centrifuge at 12000 rpm for 1 min, then add 500 μL SWB2, centrifuge at 12000 rpm for 1 min, and finally elute with 50 μL eluent to obtain a DNA solution.
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