Method for extracting DNA by using silicic bead method in combination with centrifuge shield

The invention discloses a method for extracting DNA by using a silicic bead method in combination with a centrifuge shield. The method comprises the following steps: 1) putting detected materials into an inner sleeve of the centrifuge shield, uniformly mixing a TES buffer solution, sodium dodecyl sa...

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description The invention discloses a method for extracting DNA by using a silicic bead method in combination with a centrifuge shield. The method comprises the following steps: 1) putting detected materials into an inner sleeve of the centrifuge shield, uniformly mixing a TES buffer solution, sodium dodecyl sarcosinate and proteinase K, adding the obtained mixture into the inner sleeve, and placing the obtained object on a metal bath to crack; 2) centrifugalizing the obtained object in a centrifuge, and adding an adsorption solution for re-centrifugalizing; 3) removing the inner sleeve, adding an adsorption solution and silica bead turbid liquid, and standing for adsorption; 4) centrifugalizing the obtained object so as to remove a supernatant; 5) centrifugalizing the obtained object so as to absorb raffinate; 6) adding absolute alcohol, and centrifugalizing the obtained mixture so as to remove supernatant; 7) centrifugalizing the obtained object so as to absorb raffinate; 8) drying silica beads, and adding eluant for incubating; and 9) adding the obtained product into PCR amplification liquid to amplify. The method disclosed by the invention is high in detection rate, and the final template inventory is 50-70%, therefore, the method is especially applicable to the inspection of trace detected materials, and can detect field DNA detected materials with a template content of over 100 pg.
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The method comprises the following steps: 1) putting detected materials into an inner sleeve of the centrifuge shield, uniformly mixing a TES buffer solution, sodium dodecyl sarcosinate and proteinase K, adding the obtained mixture into the inner sleeve, and placing the obtained object on a metal bath to crack; 2) centrifugalizing the obtained object in a centrifuge, and adding an adsorption solution for re-centrifugalizing; 3) removing the inner sleeve, adding an adsorption solution and silica bead turbid liquid, and standing for adsorption; 4) centrifugalizing the obtained object so as to remove a supernatant; 5) centrifugalizing the obtained object so as to absorb raffinate; 6) adding absolute alcohol, and centrifugalizing the obtained mixture so as to remove supernatant; 7) centrifugalizing the obtained object so as to absorb raffinate; 8) drying silica beads, and adding eluant for incubating; and 9) adding the obtained product into PCR amplification liquid to amplify. 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The method comprises the following steps: 1) putting detected materials into an inner sleeve of the centrifuge shield, uniformly mixing a TES buffer solution, sodium dodecyl sarcosinate and proteinase K, adding the obtained mixture into the inner sleeve, and placing the obtained object on a metal bath to crack; 2) centrifugalizing the obtained object in a centrifuge, and adding an adsorption solution for re-centrifugalizing; 3) removing the inner sleeve, adding an adsorption solution and silica bead turbid liquid, and standing for adsorption; 4) centrifugalizing the obtained object so as to remove a supernatant; 5) centrifugalizing the obtained object so as to absorb raffinate; 6) adding absolute alcohol, and centrifugalizing the obtained mixture so as to remove supernatant; 7) centrifugalizing the obtained object so as to absorb raffinate; 8) drying silica beads, and adding eluant for incubating; and 9) adding the obtained product into PCR amplification liquid to amplify. The method disclosed by the invention is high in detection rate, and the final template inventory is 50-70%, therefore, the method is especially applicable to the inspection of trace detected materials, and can detect field DNA detected materials with a template content of over 100 pg.</abstract><oa>free_for_read</oa></addata></record>
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subjects BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
title Method for extracting DNA by using silicic bead method in combination with centrifuge shield
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