{"id":137,"date":"2020-02-07T16:25:42","date_gmt":"2020-02-07T16:25:42","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/ask-a-scientist\/?p=137"},"modified":"2020-02-07T16:25:42","modified_gmt":"2020-02-07T16:25:42","slug":"what-is-qpcr","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/ask-a-scientist\/what-is-qpcr\/","title":{"rendered":"What is qPCR?"},"content":{"rendered":"<p><img loading=\"lazy\" decoding=\"async\" src=\"http:\/\/www.thermofisher.com\/blog\/ask-a-scientist\/wp-content\/uploads\/sites\/15\/2020\/02\/banner_20what20is20qpcr_-2.png\" alt=\"What is qPCR?\" width=\"600\" height=\"250\" style=\"margin-left: auto;margin-right: auto\" \/><\/p>\n<p><a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/pcr\/real-time-pcr.html?cid=corp_ask-a-scientist_blog\" target=\"_blank\" rel=\"noopener noreferrer\">qPCR stands for <strong>quantitative polymerase chain reaction<\/strong><\/a> and is a technology used for measuring DNA using PCR.&nbsp;<\/p>\n<h2>PCR vs. qPCR<\/h2>\n<p>Most applications of polymerase chain reaction focus on its utility as a way to turn a small amount of DNA into a larger amount of DNA.&nbsp;This is necessary for gel electrophoresis and most forms of DNA sequencing.&nbsp; But, it also limits how informative PCR can be. It&#8217;s&nbsp; often seen as a step in another measurement, rather than a tool on its own.<\/p>\n<p>Quantitative PCR adds two elements to the standard PCR process:<\/p>\n<ul>\n<li>Fluorescent dye<\/li>\n<li>Fluorometer<\/li>\n<\/ul>\n<p>These two elements turn qPCR from a processing step in another procedure &#8211; to a measurement technique in its own right.&nbsp;<\/p>\n<p>Thermal cyclers meant for use with qPCR include a fluorometer to detect that fluorescence.<\/p>\n<p>The fluorometer detects that fluorescence in real time as the thermal cycler runs, giving readings throughout the amplification process of the PCR.<\/p>\n<p>As a result, quantitative PCR is also called real-time PCR or RT-PCR.<\/p>\n<p>In combination with appropriate standard curves and reference values, this real-time information about reaction rates and times translates into information about relative and absolute amounts of DNA present.<\/p>\n<h2>Gene Expression with qPCR&nbsp;<\/h2>\n<p>One of the foremost strengths of qPCR is the ability to measure gene expression.<\/p>\n<p>Gene expression, or mRNA synthesis, is a critical part of protein synthesis. Gene expression is an area of active inquiry for molecular biologists &#8211; which aids in understanding numerous biological pathways and diseases.&nbsp;Using reverse transcriptase, the researcher creates complementary DNA (cDNA) copies of the RNA in their sample, and then performs qPCR with that cDNA.<\/p>\n<p>This process is called reverse-transcription polymerase chain reaction.&nbsp;<\/p>\n<p>The resulting readings provide information about the amount of&nbsp; otherwise difficult-to-measure mRNA present in the original, un-reverse-transcribed sample.<\/p>\n<p>The kind of data generated by a qPCR run depends heavily on the primers and dyes used.<\/p>\n<p>Typically, a single qPCR collects data on a single gene, since the dyes used in qPCR are usually nonspecific and will merge multiple genes into a single reading if multiple primers are used in the same mix. However, with fluorescent DNA probes rather than dyes, it is possible to &ldquo;multiplex&rdquo; qPCR and measure multiple DNA targets, as long as each target&rsquo;s corresponding probe fluoresces at a different frequency.<\/p>\n<p>For more information on the applications, theory and practice of qPCR, visit our <a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/pcr\/real-time-pcr\/real-time-pcr-learning-center\/real-time-pcr-basics\/ask-taqman-video-series.html?cid=corp_ask-a-scientist_blog\" target=\"_blank\" rel=\"noopener noreferrer\">&#8216;Ask TaqMan&#8217; resource center<\/a>.<a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/pcr\/real-time-pcr.html\"><br \/><\/a><\/p>\n<p><a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/pcr\/real-time-pcr\/real-time-pcr-learning-center\/real-time-pcr-basics\/ask-taqman-video-series.html?cid=corp_ask-a-scientist_blog\" target=\"_blank\" rel=\"noopener noreferrer\"><img loading=\"lazy\" decoding=\"async\" src=\"http:\/\/www.thermofisher.com\/blog\/ask-a-scientist\/wp-content\/uploads\/sites\/15\/2020\/02\/cta_20what20is20qpcr_-2.png\" alt=\"qPCR resource center\" width=\"560\" height=\"315\" style=\"margin-left: auto;margin-right: auto\" \/><\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>qPCR stands for quantitative polymerase chain reaction and is a technology used for measuring DNA using PCR.&nbsp; PCR vs. qPCR Most applications of polymerase chain reaction focus on its utility as a way to turn a small amount of DNA into a larger amount of DNA.&nbsp;This is necessary for gel electrophoresis and most forms of<\/p>\n","protected":false},"author":1178,"featured_media":232,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_feature_clip_id":0,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_post_was_ever_published":false},"categories":[3],"tags":[71,72],"division":[],"class_list":{"0":"post-137","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-life-sciences","8":"tag-pcr","9":"tag-qpcr","10":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO Premium plugin v27.8 (Yoast SEO v27.8) - https:\/\/yoast.com\/product\/yoast-seo-premium-wordpress\/ -->\n<title>What is qPCR? | Ask a Scientist<\/title>\n<meta name=\"description\" content=\"qPCR stands for quantitative polymerase chain reaction and is a technology for measuring DNA amounts with PCR. 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