{"id":121400,"date":"2026-08-26T15:01:20","date_gmt":"2026-08-26T15:01:20","guid":{"rendered":"https:\/\/phoeniximpact.ca\/2026\/08\/26\/quantification-de-ladn\/"},"modified":"2026-08-26T18:38:23","modified_gmt":"2026-08-26T18:38:23","slug":"quantification-de-ladn","status":"publish","type":"post","link":"https:\/\/phoeniximpact.ca\/fr\/2026\/08\/26\/quantification-de-ladn\/","title":{"rendered":"Quantification de l’ADN"},"content":{"rendered":"

[vc_row row_height_percent=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb h_padding=\u00a0\u00bb2″ top_padding=\u00a0\u00bb2″ bottom_padding=\u00a0\u00bb2″ back_color=\u00a0\u00bbcolor-366045″ overlay_alpha=\u00a0\u00bb50″ equal_height=\u00a0\u00bbyes\u00a0\u00bb gutter_size=\u00a0\u00bb3″ column_width_percent=\u00a0\u00bb100″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ uncode_shortcode_id=\u00a0\u00bb224020″ back_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb][vc_column column_width_percent=\u00a0\u00bb100″ position_vertical=\u00a0\u00bbmiddle\u00a0\u00bb align_horizontal=\u00a0\u00bbalign_center\u00a0\u00bb gutter_size=\u00a0\u00bb2″ overlay_alpha=\u00a0\u00bb50″ shift_x=\u00a0\u00bb0″ shift_y=\u00a0\u00bb0″ shift_y_down=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ medium_width=\u00a0\u00bb4″ mobile_width=\u00a0\u00bb0″ width=\u00a0\u00bb2\/3″ uncode_shortcode_id=\u00a0\u00bb122956″][vc_custom_heading text_color=\u00a0\u00bbcolor-xsdn\u00a0\u00bb heading_semantic=\u00a0\u00bbh1″ text_size=\u00a0\u00bbh4″ text_transform=\u00a0\u00bbuppercase\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb989194″ text_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb]Validation<\/span>[\/vc_custom_heading][vc_custom_heading text_color=\u00a0\u00bbcolor-xsdn\u00a0\u00bb heading_semantic=\u00a0\u00bbh1″ text_size=\u00a0\u00bbfontsize-155944″ text_weight=\u00a0\u00bb600″ text_space=\u00a0\u00bbfontspace-781688″ uncode_shortcode_id=\u00a0\u00bb373306″ text_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb]Quantification de l’ADN<\/span>[\/vc_custom_heading][vc_empty_space empty_h=\u00a0\u00bb1″][vc_button button_color=\u00a0\u00bbcolor-118531″ radius=\u00a0\u00bbbtn-round\u00a0\u00bb outline=\u00a0\u00bbyes\u00a0\u00bb border_width=\u00a0\u00bb0″ link=\u00a0\u00bburl:https%3A%2F%2Fphoeniximpact.ca%2Ffr%2Fvalidation%2F%23contact|title:Contact|\u00a0\u00bb button_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb988730″]Sugg\u00e9rer des tests suppl\u00e9mentaires[\/vc_button][\/vc_column][vc_column column_width_percent=\u00a0\u00bb100″ gutter_size=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb column_padding=\u00a0\u00bb0″ back_image=\u00a0\u00bb121423″ parallax=\u00a0\u00bbyes\u00a0\u00bb overlay_alpha=\u00a0\u00bb50″ shift_x=\u00a0\u00bb0″ shift_y=\u00a0\u00bb0″ shift_y_down=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ medium_width=\u00a0\u00bb4″ mobile_width=\u00a0\u00bb0″ width=\u00a0\u00bb1\/3″ uncode_shortcode_id=\u00a0\u00bb194376″][vc_single_image media=\u00a0\u00bb119635″ media_width_percent=\u00a0\u00bb100″ alignment=\u00a0\u00bbright\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb173810″][\/vc_column][\/vc_row][vc_row row_height_percent=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb h_padding=\u00a0\u00bb2″ top_padding=\u00a0\u00bb3″ bottom_padding=\u00a0\u00bb0″ overlay_alpha=\u00a0\u00bb50″ gutter_size=\u00a0\u00bb3″ column_width_percent=\u00a0\u00bb100″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ uncode_shortcode_id=\u00a0\u00bb146190″][vc_column column_width_percent=\u00a0\u00bb100″ align_horizontal=\u00a0\u00bbalign_center\u00a0\u00bb gutter_size=\u00a0\u00bb3″ overlay_alpha=\u00a0\u00bb50″ shift_x=\u00a0\u00bb0″ shift_y=\u00a0\u00bb0″ shift_y_down=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ medium_width=\u00a0\u00bb0″ mobile_width=\u00a0\u00bb0″ width=\u00a0\u00bb1\/1″ uncode_shortcode_id=\u00a0\u00bb926637″][vc_custom_heading text_color=\u00a0\u00bbcolor-118531″ heading_semantic=\u00a0\u00bbh3″ text_size=\u00a0\u00bbh3″ text_weight=\u00a0\u00bb700″ text_transform=\u00a0\u00bbuppercase\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb207137″ text_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb]Quantification de faibles concentrations d’ADN plasmidique et d’ADN simple brin par dosage fluorim\u00e9trique[\/vc_custom_heading][\/vc_column][\/vc_row][vc_row row_height_percent=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb h_padding=\u00a0\u00bb2″ top_padding=\u00a0\u00bb0″ bottom_padding=\u00a0\u00bb2″ overlay_alpha=\u00a0\u00bb50″ gutter_size=\u00a0\u00bb3″ column_width_percent=\u00a0\u00bb100″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ uncode_shortcode_id=\u00a0\u00bb199233″][vc_column column_width_percent=\u00a0\u00bb100″ gutter_size=\u00a0\u00bb3″ override_padding=\u00a0\u00bbyes\u00a0\u00bb column_padding=\u00a0\u00bb2″ style=\u00a0\u00bblight\u00a0\u00bb overlay_alpha=\u00a0\u00bb100″ shift_x=\u00a0\u00bb0″ shift_y=\u00a0\u00bb0″ shift_y_down=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ medium_width=\u00a0\u00bb0″ mobile_width=\u00a0\u00bb0″ width=\u00a0\u00bb1\/1″ uncode_shortcode_id=\u00a0\u00bb566008″][vc_column_text text_lead=\u00a0\u00bbyes\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb141647″]<\/p>\n

Afin de d\u00e9terminer si le reconditionnement des tubes de microcentrifugeuse affecte la pr\u00e9cision de la quantification des acides nucl\u00e9iques (dans la gamme des ng et des \u03bcg), nous avons r\u00e9alis\u00e9 des dosages fluorim\u00e9triques sur deux types d\u2019ADN : l\u2019ADN double brin (dsADN), correspondant au plasmide pCL7c (~5 kb), \u00e0 l\u2019aide du kit AccuBlue NextGen<\/span><\/a> (Biotium), et de l\u2019ADN simple brin (ssADN) \u00e0 l\u2019aide du kit Helixyte Green<\/span><\/a> (AAT Bioquest). Les analyses ont \u00e9t\u00e9 r\u00e9alis\u00e9es dans des tubes de microcentrifugeuse en polypropyl\u00e8ne de 1,5 ml pour l\u2019ADN double brin et dans des tubes de 0,6 ml pour l\u2019ADN simple brin.<\/p>\n

Les tubes de microcentrifugeuse ont \u00e9t\u00e9 soumis \u00e0 un cycle de reconditionnement, puis r\u00e9utilis\u00e9s pour r\u00e9aliser les dosages, en comparaison avec des consommables neufs : deux marques dans le cas du simple brin (M1 Invitrogen Low-binding, M2 Corning) et trois marques dans le cas du double brin (M1 Invitrogen Low-binding, M2 Corning, M3 FunChem). Les deux formats ont suivi les protocoles standard, avec 4 ou 5 \u00e9talons de concentration et un blanc au Tris-EDTA dans chaque condition.<\/p>\n

Pour l’ADN simple brin, trois ADN distincts ont \u00e9t\u00e9 test\u00e9s : Myt1 Reverse et Myt1 Forward qui sont des primers d’environ 45 paires de bases (pb), ainsi que 3XNLS-BFP-R, une s\u00e9quence d’ADN codant une prot\u00e9ine fluorescente bleue d’environ 800 \u00e0 1 100 pb. Les concentrations test\u00e9es vont de 2.3 ng\/mL \u00e0 2 \u03bcg\/mL. Les courbes standard et les graphiques pr\u00e9sent\u00e9s ici correspondent au Myt1 Forward, donn\u00e9 \u00e0 titre d’exemple repr\u00e9sentatif. Pour l\u2019ADN plasmidique, les concentrations test\u00e9es vont de 0.5 ng\/mL \u00e0 0.3 \u03bcg\/mL.<\/p>\n

Chaque condition a \u00e9t\u00e9 mesur\u00e9e en trois r\u00e9plicats. Les courbes standards, les moyennes des 3 r\u00e9plicats et les coefficients de variation (CV<14%) ont \u00e9t\u00e9 compar\u00e9s entre consommables neufs et reconditionn\u00e9s afin d’\u00e9valuer tout effet sur la pr\u00e9cision et la variabilit\u00e9 des mesures.<\/p>\n

[\/vc_column_text][\/vc_column][\/vc_row][vc_row row_height_percent=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb h_padding=\u00a0\u00bb2″ top_padding=\u00a0\u00bb0″ bottom_padding=\u00a0\u00bb0″ overlay_alpha=\u00a0\u00bb50″ gutter_size=\u00a0\u00bb3″ column_width_percent=\u00a0\u00bb100″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ content_parallax=\u00a0\u00bb0″ uncode_shortcode_id=\u00a0\u00bb158150″][vc_column width=\u00a0\u00bb1\/1″][vc_single_image media=\u00a0\u00bb121432″ media_width_percent=\u00a0\u00bb100″ uncode_shortcode_id=\u00a0\u00bb142359″][\/vc_column][\/vc_row][vc_row row_height_percent=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb h_padding=\u00a0\u00bb2″ top_padding=\u00a0\u00bb0″ bottom_padding=\u00a0\u00bb2″ overlay_alpha=\u00a0\u00bb50″ gutter_size=\u00a0\u00bb3″ column_width_percent=\u00a0\u00bb100″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ content_parallax=\u00a0\u00bb0″ uncode_shortcode_id=\u00a0\u00bb172124″][vc_column width=\u00a0\u00bb1\/1″][vc_column_text uncode_shortcode_id=\u00a0\u00bb123483″]Figure 1. Quantification de l’ADN sur des tubes neufs et des tubes reconditionn\u00e9s. Aucune diff\u00e9rence significative n’a \u00e9t\u00e9 observ\u00e9e. Chaque concentration a \u00e9t\u00e9 test\u00e9e en 3 r\u00e9plicats par condition.[\/vc_column_text][\/vc_column][\/vc_row][vc_row row_height_percent=\u00a0\u00bb0″ override_padding=\u00a0\u00bbyes\u00a0\u00bb h_padding=\u00a0\u00bb2″ top_padding=\u00a0\u00bb0″ bottom_padding=\u00a0\u00bb0″ overlay_alpha=\u00a0\u00bb50″ equal_height=\u00a0\u00bbyes\u00a0\u00bb gutter_size=\u00a0\u00bb2″ column_width_percent=\u00a0\u00bb100″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ uncode_shortcode_id=\u00a0\u00bb139950″][vc_column column_width_percent=\u00a0\u00bb100″ gutter_size=\u00a0\u00bb0″ back_color=\u00a0\u00bbcolor-xsdn\u00a0\u00bb overlay_alpha=\u00a0\u00bb50″ shift_x=\u00a0\u00bb0″ shift_y=\u00a0\u00bb-2″ shift_y_down=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ medium_width=\u00a0\u00bb0″ mobile_width=\u00a0\u00bb0″ width=\u00a0\u00bb1\/2″ uncode_shortcode_id=\u00a0\u00bb157015″ back_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb][vc_row_inner row_inner_height_percent=\u00a0\u00bb100″ overlay_color=\u00a0\u00bbcolor-xsdn\u00a0\u00bb overlay_alpha=\u00a0\u00bb30″ equal_height=\u00a0\u00bbyes\u00a0\u00bb gutter_size=\u00a0\u00bb3″ shift_y=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ limit_content=\u00a0\u00bb\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb662321″ overlay_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb][vc_column_inner column_width_percent=\u00a0\u00bb100″ gutter_size=\u00a0\u00bb1″ override_padding=\u00a0\u00bbyes\u00a0\u00bb column_padding=\u00a0\u00bb2″ style=\u00a0\u00bblight\u00a0\u00bb back_color=\u00a0\u00bbaccent\u00a0\u00bb overlay_alpha=\u00a0\u00bb50″ radius=\u00a0\u00bbxl\u00a0\u00bb shift_x=\u00a0\u00bb0″ shift_y=\u00a0\u00bb0″ shift_y_down=\u00a0\u00bb0″ z_index=\u00a0\u00bb0″ medium_width=\u00a0\u00bb4″ mobile_width=\u00a0\u00bb0″ width=\u00a0\u00bb1\/2″ mobile_height=\u00a0\u00bb300″ uncode_shortcode_id=\u00a0\u00bb184775″ back_color_type=\u00a0\u00bbuncode-palette\u00a0\u00bb][vc_icon position=\u00a0\u00bbleft\u00a0\u00bb icon_image=\u00a0\u00bb120655″ media_size=\u00a0\u00bb60″ text_size=\u00a0\u00bbh2″ align=\u00a0\u00bbleft\u00a0\u00bb title=\u00a0\u00bbR\u00e9sultats\u00a0\u00bb uncode_shortcode_id=\u00a0\u00bb888622″][\/vc_icon][vc_column_text uncode_shortcode_id=\u00a0\u00bb123483″]<\/p>\n

Les tubes reconditionn\u00e9s ont permis d’obtenir des courbes standard comparables \u00e0 celles obtenues avec des consommables neufs, et ce pour les deux types d’ADN (panneaux A et B). Aucune diff\u00e9rence marqu\u00e9e n’a \u00e9t\u00e9 d\u00e9tect\u00e9e au niveau du signal pour aucun des \u00e9talons, avec des \u00e9carts entre conditions tr\u00e8s faibles (panneaux C et D). La marque Low-binding (M1) n’a montr\u00e9 aucun avantage d\u00e9tectable aux concentrations \u00e9tudi\u00e9es.<\/p>\n

Les \u00e9carts et les coefficients de variation ont l\u00e9g\u00e8rement vari\u00e9 d’une concentration \u00e0 l’autre et d’un r\u00e9plicat \u00e0 l’autre, mais sans tendance constante en faveur de l’un ou l’autre groupe, ce qui sugg\u00e8re que la variation observ\u00e9e refl\u00e8te des facteurs exp\u00e9rimentaux normaux comme la pr\u00e9paration des dilutions, le pipetage ou le moment de la mesure, plut\u00f4t qu’un effet du reconditionnement.<\/p>\n

Aux concentrations les plus basses, la variabilit\u00e9 est domin\u00e9e par le bruit instrumental \u00e0 l’approche de la limite de d\u00e9tection des kits, et non par un ph\u00e9nom\u00e8ne d’adsorption d\u00e9pendant de la concentration.<\/p>\n

Ces r\u00e9sultats confirment que les tubes de microcentrifugation reconditionn\u00e9s peuvent \u00eatre r\u00e9utilis\u00e9s pour la quantification de l’ADN, aussi bien plasmidique que simple brin, sans perte de pr\u00e9cision ni de fiabilit\u00e9 m\u00eame pour de tr\u00e8s faibles concentrations.<\/p>\n

[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"

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