{"id":8964,"date":"2017-02-01T11:57:52","date_gmt":"2017-02-01T10:57:52","guid":{"rendered":"http:\/\/www.neb-online.fr\/en\/neb-en\/cloning-synthetic-biology\/dna-assembly\/golden-gate-assembly\/"},"modified":"2022-11-10T16:03:36","modified_gmt":"2022-11-10T15:03:36","slug":"golden-gate-assembly","status":"publish","type":"page","link":"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/golden-gate-assembly\/","title":{"rendered":"Golden Gate Assembly"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row][vc_column width=&#8221;1\/2&#8243;][vc_column_text]<\/p>\n<h2>Golden Gate Assembly<\/h2>\n<p>[\/vc_column_text][vc_column_text]The efficient and seamless assembly of DNA fragments, commonly referred to as <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/E1601@\" target=\"_blank\" rel=\"noopener noreferrer\">Golden Gate assembly<\/a>, has its origins in 1996, when for the first time it was shown that multiple inserts could be assembled into a vector backbone using only the sequential or simultaneous activities of a single type IIS <a title=\"Restriction Enzymes\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/restrictionsenzyme\/\">restriction enzyme<\/a> and <a title=\"T4 DNA Ligase\" href=\"https:\/\/www.neb-online.fr\/t4-dna-ligase\/\">T4 DNA ligase<\/a>.[\/vc_column_text][\/vc_column][vc_column width=&#8221;1\/2&#8243;]<div class=\"vbox\">\n\t\t\t\t<span class=\"vhead\">Advantages:<\/span>\n\t\t\t\t\t<ul class=\"vlist\">\n\t\t\t\t\t\t<li><span>The overhang sequence created is not dictated by the REase, and therefore no scar sequence is introduced.<br \/><\/span><\/li><li><span>The fragment-specific sequence of the overhangs allows orderly assembly of multiple fragments simultaneously.<br \/><\/span><\/li><li><span>The restriction site is eliminated from the ligated product, so digestion and ligation can be carried out simultaneously.<\/span><\/li>\n\t\t\t\t\t<\/ul>\n\t\t\t<\/div>[\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]Golden Gate Assembly and its derivative methods exploit the ability of Type IIS restriction endonucleases (REases) to cleave DNA outside of the recognition sequence. The inserts and cloning vectors are designed to place the Type IIS recognition site distal to the cleavage site, such that the Type IIS REase can remove the recognition sequence from the assembly.<\/p>\n<p>The net result is the ordered and seamless assembly of DNA fragments in one reaction. The accuracy of the assembly is dependent on the length of the overhang sequences. Therefore, Type IIS REases that create 4-base overhangs (such as <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/R0535@\" target=\"_blank\" rel=\"noopener noreferrer\">BsaI<\/a>\/<a href=\"https:\/\/shop.neb-online.fr\/en\/art\/R3733@\" target=\"_blank\" rel=\"noopener noreferrer\">BsaI-HF\u00aev2<\/a>, <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/R0539@\" target=\"_blank\" rel=\"noopener noreferrer\">BbsI<\/a>, <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/R0580@\" target=\"_blank\" rel=\"noopener noreferrer\">BsmBI, <\/a><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/R0734@\">Esp3I<\/a> and <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/R0745@\">PaqCI<\/a>) are preferred.<\/p>\n<p>Insert assembly calls for careful design of overhangs to direct the assembly, as well as verification that the Type IIS REase sites used are not present in the fragments for the assembly of the expected product. The use of web tools such as the <a href=\"https:\/\/goldengate.neb.com\/\" target=\"_blank\" rel=\"noopener noreferrer\">NEB Golden Gate Assembly Tool<\/a> greatly simplifies both processes, making Golden Gate Assembly a robust technology that assembles multiple DNA fragments, even if repetitive elements are present and can, if wished, introduce multiple site-directed mutations. Golden Gate Assembly has been widely used in the construction of custom-specific TALENs for in vivo gene editing, among other applications.[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_separator][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]<\/p>\n<h3>Golden Gate Workflow<\/h3>\n<p>[\/vc_column_text][vc_single_image image=&#8221;2037&#8243; img_size=&#8221;full&#8221;][\/vc_column][\/vc_row][vc_row][vc_column][vc_separator][vc_column_text]<\/p>\n<h2>Available Products<strong><br \/>\n<\/strong><\/h2>\n<table style=\"width: 760px;\" border=\"0\" cellpadding=\"5\">\n<tbody>\n<tr style=\"background-color: #f3b281;\">\n<th style=\"border-bottom: solid #eb7e2d;\" width=\"400\" height=\"40\">PRODUCT<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" width=\"100\" height=\"40\">REF.<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" width=\"100\">SIZE<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" colspan=\"1\" width=\"160\">PRICE<\/th>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\"><span style=\"color: #f58025;\"><strong>NEW<\/strong><\/span> NEBridge<sup>TM<\/sup> Ligase Master Mix<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">M1100S<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">50 rxns<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"10px\"><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/m1100@\"><img decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" src=\"https:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/05\/shop_icon1.gif\" alt=\"shop_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\"><span style=\"color: #f58025;\"><strong>NEW<\/strong><\/span> NEB Golden Gate Assembly Kit (BsmBI-v2)<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E1602S\/L<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">20 \/ 100 rxns<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"10px\"><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/e1602@\"><img decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" src=\"https:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/05\/shop_icon1.gif\" alt=\"shop_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\">NEB Golden Gate Assembly Mix (BsaI-HF v2)<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E1601S\/L<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">20 \/ 100 rxns<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"10px\"><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/e1601s\"><img decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" src=\"http:\/\/www.neb-online.de\/wp-content\/uploads\/2015\/05\/shop_icon1.gif\" alt=\"shop_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>[\/vc_column_text][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p>[vc_row][vc_column width=&#8221;1\/2&#8243;][vc_column_text] Golden Gate Assembly [\/vc_column_text][vc_column_text]The efficient and seamless assembly of DNA fragments, commonly referred to as Golden Gate assembly, has its origins in 1996, when for the first time it was shown that multiple inserts could be assembled into a vector backbone using only the sequential or simultaneous activities of a single type IIS&#8230;  <a class=\"excerpt-read-more\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/golden-gate-assembly\/\" title=\"Read Golden Gate Assembly\">Read more &raquo;<\/a><\/p>\n","protected":false},"author":15,"featured_media":8943,"parent":2066,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"page-sidebar-cloning-synthetic-biology.php","meta":{"footnotes":""},"class_list":["post-8964","page","type-page","status-publish","has-post-thumbnail","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.0 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Golden Gate Assembly - New England Biolabs France<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/golden-gate-assembly\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Golden Gate Assembly - New England Biolabs France\" \/>\n<meta property=\"og:description\" content=\"[vc_row][vc_column width=&#8221;1\/2&#8243;][vc_column_text] Golden Gate Assembly [\/vc_column_text][vc_column_text]The efficient and seamless assembly of DNA fragments, commonly referred to as Golden Gate assembly, has its origins in 1996, when for the first time it was shown that multiple inserts could be assembled into a vector backbone using only the sequential or simultaneous activities of a single type IIS... 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