{"id":36,"date":"2014-05-16T15:33:38","date_gmt":"2014-05-16T19:33:38","guid":{"rendered":"http:\/\/blogs.nd.edu\/bios21201\/?page_id=36"},"modified":"2014-07-12T21:14:22","modified_gmt":"2014-07-13T01:14:22","slug":"polymerase-chain-reaction-pcr","status":"publish","type":"page","link":"https:\/\/sites.nd.edu\/bios21201\/techniques\/polymerase-chain-reaction-pcr\/","title":{"rendered":"Polymerase Chain Reaction (PCR)"},"content":{"rendered":"<p style=\"color: #222222\">Polymerase chain reaction (PCR) is a powerful technique used in several areas of research.\u00a0 PCR takes advantage of the basic principles of DNA replication and allows for specific copying of a DNA sequence.<\/p>\n<p>Here is a 3-D simulation of the process:<\/p>\n<p><iframe loading=\"lazy\" title=\"Polymerase Chain Reaction (PCR)\" width=\"625\" height=\"352\" src=\"https:\/\/www.youtube.com\/embed\/2KoLnIwoZKU?feature=oembed\" frameborder=\"0\" allow=\"accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share\" referrerpolicy=\"strict-origin-when-cross-origin\" allowfullscreen><\/iframe><\/p>\n<p style=\"color: #222222\">\u00a0A PCR reaction includes 3 basic steps which are repeated for 20-30 cycles:<\/p>\n<div style=\"color: #222222\">\n<ul style=\"font-style: inherit\">\n<li style=\"font-style: inherit\">Denature:\u00a0 The two strands of a DNA molecule are split from each other by simple heating.\u00a0 Recall that the two sides of a DNA helix are held together by relatively weak hydrogen bonds between the base pairs.<\/li>\n<\/ul>\n<ul style=\"font-style: inherit\">\n<li style=\"font-style: inherit\">Anneal:\u00a0 Each PCR reaction uses a specific pair of DNA primers (short, single stranded DNA molecules).\u00a0 These primers are designed to flank the area of DNA to be copied in the reaction.\u00a0\u00a0 The reaction is cooled to a temperature (somewhere between 50-65\u00b0 C) where the DNA strands start to reattach, and the primers stick to their complementary sequence.<\/li>\n<\/ul>\n<ul style=\"font-style: inherit\">\n<li style=\"font-style: inherit\">Extend:\u00a0 The PCR reaction is heated to 72\u00b0C which is the optimum temperature for a DNA polymerase enzyme called Taq polymerase.\u00a0 Taq polymerase is derived from the hot spring bacterium Thermus aquaticus \u00a0and is able to remain stable through the denaturing step of each cycles.\u00a0 The Taq polymerase attaches to each primer and fills in the missing nucleotides in the 5\u2019 to 3\u2019 direction.<\/li>\n<\/ul>\n<\/div>\n<div style=\"color: #222222\">During each cycle of the three steps the amount of DNA present in the reaction<\/div>\n<div style=\"color: #222222\">doubles.<\/div>\n<div style=\"color: #222222\"><\/div>\n<div style=\"color: #222222\"><\/div>\n<div style=\"color: #222222\"><\/div>\n<div style=\"color: #222222\">Once you feel comfortable with the basic idea of PCR, read this technical chapter on PCR from Molecular Cloning:<\/div>\n<div style=\"color: #222222\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Polymerase chain reaction (PCR) is a powerful technique used in several areas of research.\u00a0 PCR takes advantage of the basic principles of DNA replication and allows for specific copying of a DNA sequence. Here is a 3-D simulation of the process: \u00a0A PCR reaction includes 3 basic steps which are repeated for 20-30 cycles: Denature:\u00a0 [&hellip;]<\/p>\n","protected":false},"author":1142,"featured_media":0,"parent":6,"menu_order":0,"comment_status":"closed","ping_status":"open","template":"","meta":{"footnotes":""},"class_list":["post-36","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/pages\/36","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/users\/1142"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/comments?post=36"}],"version-history":[{"count":6,"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/pages\/36\/revisions"}],"predecessor-version":[{"id":167,"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/pages\/36\/revisions\/167"}],"up":[{"embeddable":true,"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/pages\/6"}],"wp:attachment":[{"href":"https:\/\/sites.nd.edu\/bios21201\/wp-json\/wp\/v2\/media?parent=36"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}