{"id":626,"date":"2022-02-08T17:45:06","date_gmt":"2022-02-08T16:45:06","guid":{"rendered":"https:\/\/pgtb.fr\/?page_id=626"},"modified":"2022-02-08T17:48:25","modified_gmt":"2022-02-08T16:48:25","slug":"sequencage-long-read-oxford-nanopore-technologies","status":"publish","type":"page","link":"https:\/\/pgtb.fr\/en\/sequencage-long-read-oxford-nanopore-technologies\/","title":{"rendered":"S\u00e9quen\u00e7age Long Read &#8211; Oxford Nanopore Technologies"},"content":{"rendered":"<figure class=\"wp-block-image size-large\"><img data-recalc-dims=\"1\" fetchpriority=\"high\" decoding=\"async\" width=\"727\" height=\"319\" data-attachment-id=\"629\" data-permalink=\"https:\/\/pgtb.fr\/en\/ont_sequencing_animated\/\" data-orig-file=\"https:\/\/i0.wp.com\/pgtb.fr\/wp-content\/uploads\/2022\/02\/ont_sequencing_animated.gif?fit=727%2C319&amp;ssl=1\" data-orig-size=\"727,319\" data-comments-opened=\"1\" data-image-meta=\"{&quot;aperture&quot;:&quot;0&quot;,&quot;credit&quot;:&quot;&quot;,&quot;camera&quot;:&quot;&quot;,&quot;caption&quot;:&quot;&quot;,&quot;created_timestamp&quot;:&quot;0&quot;,&quot;copyright&quot;:&quot;&quot;,&quot;focal_length&quot;:&quot;0&quot;,&quot;iso&quot;:&quot;0&quot;,&quot;shutter_speed&quot;:&quot;0&quot;,&quot;title&quot;:&quot;&quot;,&quot;orientation&quot;:&quot;0&quot;}\" data-image-title=\"ont_sequencing_animated\" data-image-description=\"\" data-image-caption=\"\" data-large-file=\"https:\/\/i0.wp.com\/pgtb.fr\/wp-content\/uploads\/2022\/02\/ont_sequencing_animated.gif?fit=727%2C319&amp;ssl=1\" src=\"https:\/\/i0.wp.com\/pgtb.fr\/wp-content\/uploads\/2022\/02\/ont_sequencing_animated.gif?resize=727%2C319&#038;ssl=1\" alt=\"\" class=\"wp-image-629\" \/><\/figure>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Analyse<\/strong><\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">La premi\u00e8re sp\u00e9cificit\u00e9 du s\u00e9quen\u00e7age Long Reads d&rsquo;Oxford Nanopore technologies est le s\u00e9quen\u00e7age direct de brins natifs (ADN ou ARN). Pas de biais de PCR, ni de signal de fluorescence. De plus il est donc possible d&rsquo;avoir acc\u00e8s aux bases modifi\u00e9es (m\u00e9thylation, etc)<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">Les autres avantages sont principalement la rapidit\u00e9 de r\u00e9alisation des librairies et du s\u00e9quen\u00e7age, ainsi que l&rsquo;analyse en direct (selon l&rsquo;application).<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">D&rsquo;autre part cette technologie est toujours en constante \u00e9volution, les algorithmes de basecalling sont r\u00e9guli\u00e8rement mis \u00e0 jour, permettant ainsi d&rsquo;am\u00e9liorer d&rsquo;anciens jeux de donn\u00e9es (gratuitement) sans avoir \u00e0 s\u00e9quencer \u00e0 nouveau.<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">Nous pouvons utiliser 2 types de Flow Cells (MinION pour un maximum de donn\u00e9es ou Flongle pour amplicons, tests&#8230;)<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Les Applications<\/strong> sont tr\u00e8s vari\u00e9es :<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">WGS (assemblage de g\u00e9nomes, res\u00e9quen\u00e7age, \u00e9pig\u00e9n\u00e9tique (m\u00e9thylation), d\u00e9tection de CNV)<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">S\u00e9quencage cibl\u00e9 (amplicons, capture, Cas9)<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">M\u00e9tag\u00e9nomique totale ou cibl\u00e9e (eDNA), possibilit\u00e9 de s\u00e9quencer sur le terrain.<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">S\u00e9quen\u00e7age d&rsquo;ARN direct ou cDNA (caract\u00e9risation et quantifications des isoformes, ARNm pleine longueur)<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">La PGTB poss\u00e8de <strong>2 instruments<\/strong>, aux capacit\u00e9s diff\u00e9rentes, utilisant cette technologie :<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"> \u2666 Le GridION<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">S\u00e9quen\u00e7age au laboratoire<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">5 flow cells peuvent \u00eatre utilis\u00e9es en parall\u00e8le<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Basecalling haute pr\u00e9cision en direct<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>\u2666<\/strong> Le MinION<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Contr\u00f4l\u00e9 par un laptop, s\u00e9quen\u00e7age hors laboratoire possible.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">1 seule Flow cell<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">Basecalling faible pr\u00e9cision en direct (pour un aper\u00e7u des donn\u00e9es). Les datas sont ensuite basecall\u00e9es en haute qualit\u00e9 au laboratoire.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Acc\u00e8s<\/strong><\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">La pr\u00e9paration des librairies est accessible en mise \u00e0 disposition et en prestation. Le s\u00e9quen\u00e7age est exclusivement r\u00e9alis\u00e9 par la PGTB.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">La fourniture des r\u00e9actifs et consommables par la PGTB est obligatoire.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Echantillons<\/strong><\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">L\u2019utilisateur doit s\u2019informer \u00e0 propos des pr\u00e9-requis <a href=\"\/en\/Users\/Christophe\/Documents\/PGTB\/COMM\/SiteWeb\/lien\/\">ICI<\/a>. Les ADN, ARN et amplicons pourront \u00eatre contr\u00f4l\u00e9s par TapeStation 4200 et par Qubit avant la construction de la librairie.<strong><br><\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>En mise \u00e0 disposition\u2026<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Contr\u00f4les des librairies<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">La validation des librairies est faite par Tapestation 4200 et par Qubit.<strong><\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Validation du run<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">La PGTB ne peut garantir un r\u00e9sultat \u00e0 l\u2019utilisateur.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>R\u00e9sultats<\/strong><\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">Les r\u00e9sultats sont d\u00e9pos\u00e9s sur un serveur soumis \u00e0 un acc\u00e8s par identifiant et mot de passe r\u00e9serv\u00e9 \u00e0 l\u2019utilisateur.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>En prestation\u2026<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Contr\u00f4les des librairies<\/strong><\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">La validation des librairies est faite sur Bioanalyzer 2100 ou Tapestation 4200 et sur Qubit.<strong><\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>R\u00e9sultats<\/strong><\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">Les r\u00e9sultats sont d\u00e9pos\u00e9s sur un serveur soumis \u00e0 un acc\u00e8s par identifiant et mot de passe r\u00e9serv\u00e9 \u00e0 l\u2019utilisateur.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Applications :<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&#8211; Assemblage de g\u00e9nomes<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&#8211; D\u00e9tection de m\u00e9thylations<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&#8211; Caract\u00e9risation de CNV<\/p>\n\n\n\n<p class=\"has-text-align-justify wp-block-paragraph\">&#8211; RNA-Seq (caract\u00e9risation et quantification compl\u00e8te d\u2019isoformes, ARNm pleine longueur)<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&#8211; M\u00e9tagenomique totale ou cibl\u00e9e<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">&#8211; S\u00e9quencage sur le terrain (e-DNA) &#8211; MinION<\/p>","protected":false},"excerpt":{"rendered":"<p>Analyse La premi\u00e8re sp\u00e9cificit\u00e9 du s\u00e9quen\u00e7age Long Reads d&rsquo;Oxford Nanopore technologies est le s\u00e9quen\u00e7age direct de brins natifs (ADN ou ARN). Pas de biais de PCR, ni de signal de fluorescence. De plus il est donc possible d&rsquo;avoir acc\u00e8s aux bases modifi\u00e9es (m\u00e9thylation, etc) Les autres avantages sont principalement la rapidit\u00e9 de r\u00e9alisation des librairies<a class=\"more-link\" href=\"https:\/\/pgtb.fr\/en\/sequencage-long-read-oxford-nanopore-technologies\/\">Continue reading <span class=\"screen-reader-text\">\u00ab\u00a0S\u00e9quen\u00e7age Long Read &#8211; Oxford Nanopore Technologies\u00a0\u00bb<\/span><\/a><\/p>","protected":false},"author":216247060,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_crdt_document":"","advanced_seo_description":"","jetpack_seo_html_title":"","jetpack_seo_noindex":false,"footnotes":""},"class_list":["post-626","page","type-page","status-publish","hentry","entry"],"jetpack_likes_enabled":true,"jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/PdFmHm-a6","jetpack-related-posts":[],"_links":{"self":[{"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/pages\/626","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/users\/216247060"}],"replies":[{"embeddable":true,"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/comments?post=626"}],"version-history":[{"count":4,"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/pages\/626\/revisions"}],"predecessor-version":[{"id":633,"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/pages\/626\/revisions\/633"}],"wp:attachment":[{"href":"https:\/\/pgtb.fr\/en\/wp-json\/wp\/v2\/media?parent=626"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}