A method for rapidly extracting genomic DNA from peripheral blood
An extraction method and genome technology, applied in the field of rapid extraction of genomic DNA from peripheral blood, can solve the problems of cumbersome operation, increased cost and time, and achieve the effect of simple and clear steps, low reagent consumption and short time consumption
Pending Publication Date: 2019-01-15
南通中科医学检验实验室有限公司
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Problems solved by technology
In order to increase the extraction amount and purity of DNA, the operation is cumbersome, cost and time will increase
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Embodiment 1
[0026] Blood samples from 12 individuals were collected using phlebotomy;
[0027] Follow the steps below to extract DNA:
[0028] 1) Take 100 μL blood sample, add 200 μL Lysis Buffer and 20 μL Proteinase K premixed solution, and mix well by inversion. Place at 60°C for 10 minutes, and at 95°C for 10 minutes, and mix by inverting several times during this period.
[0029] 2) After standing at room temperature for 2 minutes, centrifuge at 12000 rpm for 2 minutes, and take 50 μL of supernatant.
[0030] 3) Add 90 μL of magnetic beads to the PCR tube containing the above supernatant, gently pipette and mix 6 times, incubate at room temperature for 5 minutes, and place the PCR tube on the magnetic stand for 3 minutes.
[0031] 4) Remove the supernatant, continue to place the PCR tube on the magnetic stand, add 300 μL of 75% ethanol solution to the PCR tube, and let it stand for 30 seconds.
[0032] 5) Remove the supernatant, then add 300 μL of 75% ethanol solution to the PCR tu...
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A method for rapidly extracting genomic DNA from peripheral blood comprises that follow steps: 1) adding a premix solution of 200-250 [mu]L of a lysis buffer and 20-35 [mu]L of proteinase K into 100-150 [mu]L of a blood sample, reversing and fully mixing the mixture, allowing the mixture to stand at 60 DEG C for 10 min, allowing the mixture to stand at 95 DEG C for 10 min, with the mixture being reversed and uniformly mixed in the period; 2) after the mixture is allowed to stand at room temperature for 2-5 min, performing centrifugation at 12000 rpm for 2 min, and taking 50 [mu]L of that supernatant; 3) adding 90-92 [mu]L of magnetic beads into the PCR tube equipped with the supernatant, gently blowing and fully mixing the mixture for 6 times, and incubating the mixture at room temperaturefor 5-10 min, and placing the PCR tube on a magnetic rack for 3-5min. Beneficial effects of the method are that 1) that invention only relate to three kinds of reagent in the whole DNA extraction process, and the reagent usage quantity and the sample demand quantity are low, so the cost is low; 2) that whole extraction process is short in time, each step is simple and definite, and has little influence on the amplification effect of PCR; 3) the DNA extracted by the method accord with the majority of PCR reactions and the detection of MassARRAY platform through the verification of actual items.
Description
technical field [0001] The invention relates to a rapid extraction method of peripheral blood genome DNA, which belongs to the field of biotechnology. Background technique [0002] At present, the conventional methods of peripheral blood genomic DNA extraction include phenol / chloroform method and potassium iodide method. The OD260 / OD280 of DNA extraction quality by phenol / chloroform method is about 1.97, and the DNA extraction quality OD260 / OD280 of potassium iodide method is about 2.30. In order to increase the extraction amount and purity of DNA, the operation is cumbersome, and the cost and time will increase. [0003] With the continuous development of detection technology, the sensitivity of the instrument itself has increased, the cost is low, the operation is simple, the time-consuming is short, and the obtained DNA can conform to most platforms, which has become the first choice of most genetic testing industry. Contents of the invention [0004] The purpose of th...
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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 陆军姜昕刘慧君董新波
Owner 南通中科医学检验实验室有限公司