{"id":676,"date":"2012-11-08T16:46:50","date_gmt":"2012-11-08T15:46:50","guid":{"rendered":"http:\/\/saber.comule.com\/?p=676"},"modified":"2020-11-12T19:01:07","modified_gmt":"2020-11-12T18:01:07","slug":"race-pcr","status":"publish","type":"post","link":"http:\/\/saber-benmimoun.com\/?p=676","title":{"rendered":"RACE PCR"},"content":{"rendered":"<p><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/Sans-titre.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignright size-thumbnail wp-image-679\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/Sans-titre-150x150.jpg\" alt=\"\" width=\"150\" height=\"150\" \/><\/a>On connait la s\u00e9quence codante du g\u00e8ne \u00e9tudier , donc de l&#8217;ATG au codon stop.<\/p>\n<p>on ne connait pas les extr\u00e9mit\u00e9s 5&#8217;UTR et 3&#8217;UTR de l&#8217;ARNm<\/p>\n<p>&nbsp;<\/p>\n<p><span style=\"color: #993300;\"><strong>5&#8242; RACE PCR:<\/strong><\/span><\/p>\n<p><strong>but =<\/strong> amplifier uniquement la r\u00e9gion \u00a05&#8217;UTR de l&#8217;ARNm, en passant par l&#8217;ADNc. cela permet ensuite de ne s\u00e9quencer que cette r\u00e9gion au lieu de tout l&#8217;ADNc.<\/p>\n<p><strong>Etapes:<\/strong><\/p>\n<ul>\n<li>extraction des ARNm totaux<\/li>\n<li>Ligation sp\u00e9ci\ufb01que en 5&#8242; de l ARNm\u00a0d&#8217;un oligonucl\u00e9otide simple brin (exp. polydC)<\/li>\n<li>retro-transcription en ADNc grace \u00e0 des amorces polydT<\/li>\n<li>hydrolyse de l&#8217;ARN par une RNase<\/li>\n<li>PCR avec deux amorces\n<ul>\n<li>5&#8242;&#8211;&gt;3&#8242; \u00a0\u00a0oligodG<\/li>\n<li>3&#8242;&#8211;&gt;5&#8242; \u00a0\u00a0Gene Specific primer GSP au niveau de la s\u00e9quence codante connue<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<p><span style=\"color: #993300;\"><strong>3&#8242; RACE PCR:<\/strong><\/span><\/p>\n<p><strong>but =<\/strong> amplifier uniquement la r\u00e9gion \u00a03&#8217;UTR de l&#8217;ARNm, en passant par l&#8217;ADNc. cela permet ensuite de ne s\u00e9quencer que cette r\u00e9gion au lieu de tout l&#8217;ADNc.<\/p>\n<p><strong>Etapes:<\/strong><\/p>\n<ul>\n<li>extraction des ARNm totaux<\/li>\n<li>Ligation sp\u00e9ci\ufb01que en 5&#8242; de l ARNm\u00a0d&#8217;un oligonucl\u00e9otide simple brin (exp. polydC)<\/li>\n<li>retro-transcription en ADNc grace \u00e0 des amorces polydT<\/li>\n<li>hydrolyse de l&#8217;ARN par une RNase<\/li>\n<li>PCR avec deux amorces\n<ul>\n<li>5&#8242;&#8211;&gt;3&#8242; \u00a0 Gene Specific primer GSP au niveau de la s\u00e9quence codante connue<\/li>\n<li>3&#8242;&#8211;&gt;5&#8242; \u00a0 oligodT<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>On connait la s\u00e9quence codante du g\u00e8ne \u00e9tudier , donc de l&#8217;ATG au codon stop. on ne connait pas les extr\u00e9mit\u00e9s 5&#8217;UTR et 3&#8217;UTR de l&#8217;ARNm &nbsp; 5&#8242; RACE PCR: but = amplifier uniquement la r\u00e9gion \u00a05&#8217;UTR de l&#8217;ARNm, en passant par l&#8217;ADNc. cela permet ensuite de ne s\u00e9quencer que cette r\u00e9gion au lieu de&hellip;<\/p>\n","protected":false},"author":1,"featured_media":679,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[7],"tags":[],"class_list":["post-676","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-biologie","category-7","description-off"],"_links":{"self":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/676","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=676"}],"version-history":[{"count":4,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/676\/revisions"}],"predecessor-version":[{"id":2954,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/676\/revisions\/2954"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/media\/679"}],"wp:attachment":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=676"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=676"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=676"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}