{"id":652,"date":"2012-11-03T13:06:02","date_gmt":"2012-11-03T12:06:02","guid":{"rendered":"http:\/\/saber.comule.com\/?p=652"},"modified":"2020-11-12T19:01:07","modified_gmt":"2020-11-12T18:01:07","slug":"la-demarche-transcriptome","status":"publish","type":"post","link":"http:\/\/saber-benmimoun.com\/?p=652","title":{"rendered":"La d\u00e9marche transcriptome"},"content":{"rendered":"<p><strong>\u00a0But:<\/strong> d\u00e9terminer le profil de transcription des g\u00e8ne d&#8217;un organisme donn\u00e9 dans une condition donn\u00e9e, ou comparer le profil de transcription d&#8217;un meme organisme dans diff\u00e9rentes conditions et diterminer ainsi les g\u00e8nes dont la transcription est li\u00e9 \u00e0 une condition donn\u00e9e.<\/p>\n<p><strong>Etapes:<\/strong><\/p>\n<ul>\n<li>Pr\u00e9l\u00e8vement d&#8217;un \u00e9chantillon, ou culture, de cellules en condition A et d&#8217;autres en condition B<\/li>\n<li>Extraction et purification des ARNm grace \u00e0 une colonne polydT<\/li>\n<li>RT-PCR, avec des amorces polydT et des dNTP fluorescents &#8211;&gt; synth\u00e8se ADNc marqu\u00e9e: conditionA avec Cy5 (rouge) et conditionB avec Cy3 (vert)<\/li>\n<li>D\u00e9naturation: s\u00e9paration de\u00a0 l&#8217;ARNm et de l&#8217;ADNc*<\/li>\n<li>Ajout d&#8217;une RNase pour d\u00e9grader l&#8217;ARNm<\/li>\n<li>on m\u00e9lange la m\u00eame quantit\u00e9 d&#8217;ADNc* conditionA avec l&#8217;ADNc* conditionB<\/li>\n<li>Hybridation: on d\u00e9pose le m\u00e9lange d&#8217;ADNc* sur la lame de DNA-microarrays<\/li>\n<li>Lavage<\/li>\n<li>\u00a0Exposition au UV et analyse de la fluorescence \u00e9mise:\n<ul>\n<li>les taches de couleur rouge correspondent au g\u00e8nes qui sont transcrits uniquement en conditionA.<\/li>\n<li>Les taches verte au g\u00e8nes qui\u00a0sont transcrits seulement en conditionB.<\/li>\n<li>Les taches jaunes, c&#8217;est les g\u00e8nes qui sont transcrits dans les deux cas.<\/li>\n<li>Les taches noires, c&#8217;est les g\u00e8nes qui ne sont pas transcrits dans les deux conditions.<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p><strong>Applications:<\/strong><\/p>\n<ul>\n<li>\u00c9tudier l&#8217;effet d&#8217;une drogue sur l&#8217;expression g\u00e9nique (pharmacologie, fungicides, pesticides..)<\/li>\n<li>Comparer deux conditions sp\u00e9cifiques (tissu malade vs tissu sain): applications au cancer par exemple<\/li>\n<li>\u00c9tudier des effets environnementaux<\/li>\n<li>Comprendre un programme cellulaire ou d\u00e9veloppemental<\/li>\n<li>D\u00e9crypter un r\u00e9seau de r\u00e9gulation<\/li>\n<li>STABILITE DES ARNm<\/li>\n<li>AIDE \u00c0 L&#8217;ANNOTATION DU GENOME.<\/li>\n<li>STRUCTURE DES GENES ET DES EXONS<\/li>\n<li>CARTOGRAPHIES GLOBALES TRANSCRIPTIONNELLES<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<p><strong>Remarques:<\/strong> chez les procaryotes, vu que les ARNm n&#8217;ont pas de queue polyA, on utilise des amorces al\u00e9atoire pour la PCR (donc le marquage sera par Random priming).<\/p>\n<p><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/microarray_technology.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter size-thumbnail wp-image-654\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/microarray_technology-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/p>\n<p style=\"text-align: center;\"><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/GNCHIP1.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-thumbnail wp-image-656 alignleft\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/GNCHIP1-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/GNCHIP2.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-thumbnail wp-image-657 alignleft\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/GNCHIP2-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/p>\n<p style=\"text-align: center;\"><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/GNCHIP3.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-thumbnail wp-image-658 alignleft\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/11\/GNCHIP3-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>\u00a0But: d\u00e9terminer le profil de transcription des g\u00e8ne d&#8217;un organisme donn\u00e9 dans une condition donn\u00e9e, ou comparer le profil de transcription d&#8217;un meme organisme dans diff\u00e9rentes conditions et diterminer ainsi les g\u00e8nes dont la transcription est li\u00e9 \u00e0 une condition donn\u00e9e. Etapes: Pr\u00e9l\u00e8vement d&#8217;un \u00e9chantillon, ou culture, de cellules en condition A et d&#8217;autres en&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-652","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\/652","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=652"}],"version-history":[{"count":10,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/652\/revisions"}],"predecessor-version":[{"id":2955,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/652\/revisions\/2955"}],"wp:attachment":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=652"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=652"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=652"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}