Method for rapidly extracting DNA from whole blood
An extraction method and technology for blood samples, applied in the field of rapid extraction of DNA from a large number of whole blood, can solve the problems of incompatibility with third-generation sequencing, cumbersome DNA process, long time consumption, etc., and achieve reduction of cleaning and centrifugation steps, simple processing technology, and short time consumption. Effect
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Embodiment 1
[0039] The present embodiment provides a method for rapid extraction of DNA from whole blood, and the specific steps include:
[0040] 1) Take 7.5ml of human blood sample and thaw it completely at room temperature;
[0041] 2) Centrifuge at 15000rpm at 4°C for 15min, remove the supernatant completely, leaving only the sediment at the bottom of the tube;
[0042] 3) Add 2V of 37°C preheated lysate (0.8M guanidine HCl, 30mM Tris-Cl (pH8.0), 30mM EDTA (pH8.0), 5% Tween-20, 0.5% Triton-100, 0.5% SDS, 0.5mg / ml Proteinse K), use a pipette to thoroughly break up the precipitate and mix it to a clear state;
[0043] 4) Place in a water bath at 56°C for 1 hour, and gently invert and mix once every 10 minutes;
[0044] 5) After cooling to room temperature, add an equal volume of chloroform isoamyl alcohol (24:1) for extraction once;
[0045] 6) Centrifuge at 13000rpm for 10min (16°C), and take the supernatant;
[0046] 7) Add 0.7V frozen isopropanol and 0.1V NaAc (pH5.2), mix well; ...
Embodiment 2
[0057] Take 6ml of monkey blood sample and put it in a 37°C water bath to thaw completely;
[0058] 2) Centrifuge at 15000rpm at 4°C for 15min, remove the supernatant completely, leaving only the sediment at the bottom of the tube;
[0059] 3) Add 2V of 37°C preheated lysate (0.8M guanidine HCl, 30mM Tris-Cl (pH8.0), 30mM EDTA (pH8.0), 5% Tween-20, 0.5% Triton-100, 0.5% SDS, 0.5mg / ml Proteinse K), use a pipette to thoroughly break up the precipitate and mix it to a clear state;
[0060] 4) Place in a water bath at 56°C for 1 hour, and gently invert and mix once every 10 minutes;
[0061] 5) After cooling to room temperature, add an equal volume of chloroform isoamyl alcohol (24:1) for extraction once;
[0062] 6) Centrifuge at 13000rpm for 10min (16°C), and take the supernatant;
[0063] 7) Add 0.7V frozen isopropanol and 0.1V NaAc (pH5.2), mix well;
[0064] 8) Pick out the flocculent precipitate and wash it 3 times with 75% frozen ethanol;
[0065] 9) Dry, add 300ul TEN...
Embodiment 3
[0075] The present embodiment provides a method for rapid extraction of DNA from whole blood, and the specific steps include:
[0076] 1) Take 5ml of human blood sample and thaw it completely at room temperature;
[0077] 2) Centrifuge at 15000rpm at 4°C for 15min, remove the supernatant completely, leaving only the sediment at the bottom of the tube;
[0078] 3) Add 2.5V preheated lysate (1.2M guanidine HCl, 100mM Tris-Cl (pH8.0), 80mM EDTA (pH8.0), 4% Tween-20, 0.3% Triton-100, 0.3 %SDS, 0.3mg / ml Proteinse K), the precipitate was thoroughly dispersed and mixed to a clear state with a pipette;
[0079] 4) Place in a 50°C water bath for 50 minutes, and gently invert and mix once every 5 minutes;
[0080] 5) After cooling to room temperature, add an equal volume of chloroform isoamyl alcohol (25:1) for extraction once;
[0081] 6) Centrifuge at 16000rpm for 8min (10°C), and take the supernatant;
[0082] 7) Add 1V frozen isopropanol and 0.1V NaAc (pH5.2), mix well;
[0083...
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