{"id":647,"date":"2012-11-02T18:53:41","date_gmt":"2012-11-02T17:53:41","guid":{"rendered":"http:\/\/saber.comule.com\/?p=647"},"modified":"2020-11-12T19:01:07","modified_gmt":"2020-11-12T18:01:07","slug":"pcr-dgge-denaturing-gradient-gel-electrophoresis","status":"publish","type":"post","link":"http:\/\/saber-benmimoun.com\/?p=647","title":{"rendered":"PCR-DGGE (Denaturing Gradient Gel Electrophoresis)"},"content":{"rendered":"<p>But: S\u00e9parer des mol\u00e9cules d&#8217;ADN ou d&#8217;ARN selon leur composition en CG.<\/p>\n<p>Avantages:<\/p>\n<ul>\n<li>Permet de s\u00e9parer des fragments d&#8217;ADN de tres grande taille, voir des chromosomes entiers, ce qui n&#8217;est pas possible par une \u00e9lectrophor\u00e8se classique.<\/li>\n<li>Permet de s\u00e9parer des mol\u00e9cule d&#8217;ADN ou ARN ayant la m\u00eame taille, mais ayant une composition diff\u00e9rente en GC vu leur appartenance \u00e0 des organismes diff\u00e9rents.<\/li>\n<\/ul>\n<p>Principe : Lors de leurs migration, \u00e0 travers le gradient d\u00e9naturant croissant, et lorsqu&#8217;une mol\u00e9cule atteint son point de fusion, elle devient simple brin ce qui freine son mouvement dans le gel. le CG-clamp permet d&#8217;\u00e9viter une d\u00e9naturation totale des mol\u00e9cules.<\/p>\n<p>\u00c9tapes:<a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/pic010.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-thumbnail wp-image-648 alignright\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/pic010-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/p>\n<ul>\n<li>Extraction de l&#8217;ADN de l&#8217;\u00e9chantillon<\/li>\n<li>PCR de la r\u00e9gion cible (exp. g\u00e8ne codant l&#8217;ARN 16S) et ajout d&#8217;un GC-clamp grace \u00e0 des amorces sp\u00e9ciales.<\/li>\n<li>d\u00e9p\u00f4t sur gel \u00e0 gradient d\u00e9naturant<\/li>\n<li>migration gr\u00e2ce \u00e0 un champ \u00e9lectrique<\/li>\n<li>R\u00e9v\u00e9lation et analyse du gel<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/Schema_1.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-thumbnail wp-image-650 alignright\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/Schema_1-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>But: S\u00e9parer des mol\u00e9cules d&#8217;ADN ou d&#8217;ARN selon leur composition en CG. Avantages: Permet de s\u00e9parer des fragments d&#8217;ADN de tres grande taille, voir des chromosomes entiers, ce qui n&#8217;est pas possible par une \u00e9lectrophor\u00e8se classique. Permet de s\u00e9parer des mol\u00e9cule d&#8217;ADN ou ARN ayant la m\u00eame taille, mais ayant une composition diff\u00e9rente en GC&hellip;<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[7],"tags":[],"class_list":["post-647","post","type-post","status-publish","format-standard","hentry","category-biologie","category-7","description-off"],"_links":{"self":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/647","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=647"}],"version-history":[{"count":4,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/647\/revisions"}],"predecessor-version":[{"id":2956,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/647\/revisions\/2956"}],"wp:attachment":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=647"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=647"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=647"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}