{"id":10793,"date":"2023-10-23T14:41:41","date_gmt":"2023-10-23T12:41:41","guid":{"rendered":"https:\/\/inci.neuro.unistra.fr\/?page_id=10793"},"modified":"2023-12-20T13:18:10","modified_gmt":"2023-12-20T12:18:10","slug":"quantitative-pcr-in-real-time","status":"publish","type":"page","link":"https:\/\/inci.neuro.unistra.fr\/?page_id=10793&lang=en","title":{"rendered":"Quantitative PCR in real time"},"content":{"rendered":"\t\t
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Quantitative PCR in real time<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t

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P<\/b>rinciple and objective<\/span><\/div>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t
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Quantitative PCR (real-time PCR, QPCR)<\/em> uses the basic principle of classic PCR (cyclical amplification of a DNA fragment based on an enzymological reaction)<\/em> with the difference being an amplification measured not at the end but throughout<\/span><\/span> of the reaction, and therefore in real time.<\/span><\/span> <\/span><\/p>

At each amplification cycle, the quantity of DNA is measured using a fluorescent marker such as \u201csyber green\u201d whose emission is directly proportional to the quantity of amplicons produced.<\/span><\/span> This makes it possible to obtain kinetics of the reaction and therefore quantification of the DNA whereas conventional PCR only gives the final measurement.<\/span><\/span> This technique therefore makes it possible to compare the differences in gene expression according to the experimental parameters defined by the researcher.<\/span><\/span><\/span><\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t

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E<\/b>quipment<\/span><\/div>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t
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\"\"<\/a>The team has a Biorad brand MyIQ thermocycler for 96-well plates coupled with a UV lamp and a spectrophotometer, all controlled by computer using BioRad IQ5 software.<\/span><\/span> thermocycler Mylq.<\/span><\/span> <\/span><\/p>

This equipment is complemented by a TECAN brand \u201cfreedom evo\u201d pipetting robot which allows 96-well PCR plates to be prepared in record time (approximately 7 minutes)<\/em> while avoiding pipetting errors or contamination linked to the<\/span><\/span> user.<\/span><\/span> This pipetting robot is particularly interesting if the number of plates to be made is large.<\/span><\/span> This robot is controlled via TECAN\u2019s Evoware software.<\/span><\/span><\/span><\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t

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C<\/b>ontact<\/span><\/div>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t
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\"\"The quantitative PCR technical platform is located on the 4th floor of the Institute (room 4312a)<\/em>.<\/span><\/span> <\/span><\/p>

Its mission is to make QPCR technology available to INCI researchers and staff.<\/span><\/span><\/span><\/p>

Scientific support<\/strong>: Vincent LELIEVRE<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t

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Quantitative PCR in real time Quantitative PCR (real-time PCR, QPCR) uses the basic principle of classic PCR (cyclical amplification of a DNA fragment based on an enzymological reaction) with the difference being an amplification measured not at the end but throughout of the reaction, and therefore in real time. At each amplification cycle, the quantity […]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":10781,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"ocean_post_layout":"","ocean_both_sidebars_style":"","ocean_both_sidebars_content_width":0,"ocean_both_sidebars_sidebars_width":0,"ocean_sidebar":"0","ocean_second_sidebar":"0","ocean_disable_margins":"enable","ocean_add_body_class":"","ocean_shortcode_before_top_bar":"","ocean_shortcode_after_top_bar":"","ocean_shortcode_before_header":"","ocean_shortcode_after_header":"","ocean_has_shortcode":"","ocean_shortcode_after_title":"","ocean_shortcode_before_footer_widgets":"","ocean_shortcode_after_footer_widgets":"","ocean_shortcode_before_footer_bottom":"","ocean_shortcode_after_footer_bottom":"","ocean_display_top_bar":"default","ocean_display_header":"default","ocean_header_style":"","ocean_center_header_left_menu":"0","ocean_custom_header_template":"0","ocean_custom_logo":0,"ocean_custom_retina_logo":0,"ocean_custom_logo_max_width":0,"ocean_custom_logo_tablet_max_width":0,"ocean_custom_logo_mobile_max_width":0,"ocean_custom_logo_max_height":0,"ocean_custom_logo_tablet_max_height":0,"ocean_custom_logo_mobile_max_height":0,"ocean_header_custom_menu":"0","ocean_menu_typo_font_family":"0","ocean_menu_typo_font_subset":"","ocean_menu_typo_font_size":0,"ocean_menu_typo_font_size_tablet":0,"ocean_menu_typo_font_size_mobile":0,"ocean_menu_typo_font_size_unit":"px","ocean_menu_typo_font_weight":"","ocean_menu_typo_font_weight_tablet":"","ocean_menu_typo_font_weight_mobile":"","ocean_menu_typo_transform":"","ocean_menu_typo_transform_tablet":"","ocean_menu_typo_transform_mobile":"","ocean_menu_typo_line_height":0,"ocean_menu_typo_line_height_tablet":0,"ocean_menu_typo_line_height_mobile":0,"ocean_menu_typo_line_height_unit":"","ocean_menu_typo_spacing":0,"ocean_menu_typo_spacing_tablet":0,"ocean_menu_typo_spacing_mobile":0,"ocean_menu_typo_spacing_unit":"","ocean_menu_link_color":"","ocean_menu_link_color_hover":"","ocean_menu_link_color_active":"","ocean_menu_link_background":"","ocean_menu_link_hover_background":"","ocean_menu_link_active_background":"","ocean_menu_social_links_bg":"","ocean_menu_social_hover_links_bg":"","ocean_menu_social_links_color":"","ocean_menu_social_hover_links_color":"","ocean_disable_title":"default","ocean_disable_heading":"default","ocean_post_title":"","ocean_post_subheading":"","ocean_post_title_style":"","ocean_post_title_background_color":"","ocean_post_title_background":0,"ocean_post_title_bg_image_position":"","ocean_post_title_bg_image_attachment":"","ocean_post_title_bg_image_repeat":"","ocean_post_title_bg_image_size":"","ocean_post_title_height":0,"ocean_post_title_bg_overlay":0.5,"ocean_post_title_bg_overlay_color":"","ocean_disable_breadcrumbs":"default","ocean_breadcrumbs_color":"","ocean_breadcrumbs_separator_color":"","ocean_breadcrumbs_links_color":"","ocean_breadcrumbs_links_hover_color":"","ocean_display_footer_widgets":"default","ocean_display_footer_bottom":"default","ocean_custom_footer_template":"0","omw_enable_modal_window":"enable","footnotes":""},"class_list":["post-10793","page","type-page","status-publish","hentry","entry"],"yoast_head":"\nQuantitative PCR in real time - 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