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. 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.</description><language>eng</language><subject>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</subject><creationdate>2015</creationdate><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://worldwide.espacenet.com/publicationDetails/biblio?FT=D&date=20150812&DB=EPODOC&CC=CN&NR=104830834A$$EHTML$$P50$$Gepo$$Hfree_for_read</linktohtml><link.rule.ids>230,308,780,885,25563,76318</link.rule.ids><linktorsrc>$$Uhttps://worldwide.espacenet.com/publicationDetails/biblio?FT=D&date=20150812&DB=EPODOC&CC=CN&NR=104830834A$$EView_record_in_European_Patent_Office$$FView_record_in_$$GEuropean_Patent_Office$$Hfree_for_read</linktorsrc></links><search><creatorcontrib>QI JIELI</creatorcontrib><title>Method for extracting DNA by using silicic bead method in combination with centrifuge shield</title><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.</description><subject>BEER</subject><subject>BIOCHEMISTRY</subject><subject>CHEMISTRY</subject><subject>COMPOSITIONS THEREOF</subject><subject>CULTURE MEDIA</subject><subject>ENZYMOLOGY</subject><subject>METALLURGY</subject><subject>MICROBIOLOGY</subject><subject>MICROORGANISMS OR ENZYMES</subject><subject>MUTATION OR GENETIC ENGINEERING</subject><subject>PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS</subject><subject>SPIRITS</subject><subject>VINEGAR</subject><subject>WINE</subject><fulltext>true</fulltext><rsrctype>patent</rsrctype><creationdate>2015</creationdate><recordtype>patent</recordtype><sourceid>EVB</sourceid><recordid>eNqNi0sKwjAUALNxIeodngcQKumi21IVN3blUij5vDYP0qQkr6i3V9EDuBoGZpbidkF20UIfE-CDkzJMYYBDW4N-wpw_ksmTIQMalYXx21MAE0dNQTHFAHdiBwYDJ-rnASE7Qm_XYtErn3Hz40psT8drc97hFDvMk3ofyF3T7ouykkUly1r-07wAWLE7Tg</recordid><startdate>20150812</startdate><enddate>20150812</enddate><creator>QI JIELI</creator><scope>EVB</scope></search><sort><creationdate>20150812</creationdate><title>Method for extracting DNA by using silicic bead method in combination with centrifuge shield</title><author>QI JIELI</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-epo_espacenet_CN104830834A3</frbrgroupid><rsrctype>patents</rsrctype><prefilter>patents</prefilter><language>eng</language><creationdate>2015</creationdate><topic>BEER</topic><topic>BIOCHEMISTRY</topic><topic>CHEMISTRY</topic><topic>COMPOSITIONS THEREOF</topic><topic>CULTURE MEDIA</topic><topic>ENZYMOLOGY</topic><topic>METALLURGY</topic><topic>MICROBIOLOGY</topic><topic>MICROORGANISMS OR ENZYMES</topic><topic>MUTATION OR GENETIC ENGINEERING</topic><topic>PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS</topic><topic>SPIRITS</topic><topic>VINEGAR</topic><topic>WINE</topic><toplevel>online_resources</toplevel><creatorcontrib>QI JIELI</creatorcontrib><collection>esp@cenet</collection></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>QI JIELI</au><format>patent</format><genre>patent</genre><ristype>GEN</ristype><title>Method for extracting DNA by using silicic bead method in combination with centrifuge shield</title><date>2015-08-12</date><risdate>2015</risdate><abstract>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.</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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