{"id":8955,"date":"2017-02-01T11:03:43","date_gmt":"2017-02-01T10:03:43","guid":{"rendered":"http:\/\/www.neb-online.fr\/en\/neb-en\/cloning-synthetic-biology\/dna-assembly\/nebuilder-hifi-dna-assembly\/"},"modified":"2022-08-03T16:19:05","modified_gmt":"2022-08-03T14:19:05","slug":"nebuilder-hifi-dna-assembly","status":"publish","type":"page","link":"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/nebuilder-hifi-dna-assembly\/","title":{"rendered":"NEBuilder HiFi DNA Assembly"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row][vc_column][vc_column_text]NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5\u00b4 and 3\u00b4 restriction enzyme mismatches.<\/p>\n<p>This flexible kit enables simple and fast Seamless Cloning utilizing a new proprietary high-fidelity polymerase.<\/p>\n<p>Find out why NEBuilder HiFi is the next generation of DNA assembly and cloning.[\/vc_column_text][vc_separator][vc_column_text]<\/p>\n<h2>NEBuilder HiFi DNA Assembly Workflow<\/h2>\n<p>[\/vc_column_text][vc_row_inner css=&#8221;.vc_custom_1503584886228{margin-right: 0px !important;margin-left: 0px !important;}&#8221;][vc_column_inner width=&#8221;1\/4&#8243;][vc_column_text]<strong>Illustration NEBuilder Method<\/strong><\/p>\n<p><span class=\"bildunterschrift\">Click image to enlarge<\/span>[\/vc_column_text][vc_column_text css=&#8221;.vc_custom_1466761959565{padding-top: 50px !important;}&#8221;]<strong>Illustration NEBuilder Workflow<\/strong><\/p>\n<p><span class=\"bildunterschrift\">Click image to enlarge<\/span>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=&#8221;1\/4&#8243;][vc_single_image image=&#8221;1528&#8243; onclick=&#8221;img_link_large&#8221;][vc_single_image image=&#8221;8949&#8243; onclick=&#8221;img_link_large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243; css=&#8221;.vc_custom_1466761944557{border-top-width: 2px !important;border-bottom-width: 2px !important;padding-top: 20px !important;padding-bottom: 20px !important;background-color: #f4f1e7 !important;border-top-color: #eb7e2d !important;border-top-style: solid !important;border-bottom-color: #eb7e2d !important;border-bottom-style: solid !important;}&#8221;][vc_column_text]<strong>Benefits over Gibson Assembly\u00ae Master Mix<\/strong><\/p>\n<ul>\n<li>Enjoy less screening\/re-sequencing of constructs, with virtually error-free, high-fidelity assembly<\/li>\n<li>Join DNA fragments together more efficiently, even with larger fragments or low DNA inputs<\/li>\n<li>Use NEBuilder HiFi in successive rounds of assembly, because it removes 5\u00b4- and 3\u00b4- restriction enzyme mismatches<\/li>\n<li>Bridge two double-stranded fragments with a synthetic single-stranded DNA oligo for simple and fast construction (e.g. linker insertion or gRNA libraries)<\/li>\n<li>Switch from other systems easily, because NEBuilder HiFi is compatible with Gibson Assembly-designed (and other) fragments<\/li>\n<li>Save time with simple and fast Seamless Cloning<\/li>\n<li>No licensing fee requirements from NEB for the NEBuilder products<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_column_text][\/vc_column_text][vc_separator color=&#8221;black&#8221;][\/vc_column][\/vc_row][vc_row css=&#8221;.vc_custom_1461849499284{margin-right: -1px !important;margin-left: -1px !important;border-top-width: 2px !important;border-right-width: 2px !important;border-bottom-width: 2px !important;border-left-width: 2px !important;padding-top: 20px !important;padding-bottom: 20px !important;background-color: #f4f1e7 !important;border-left-color: #eb7e2d !important;border-left-style: solid !important;border-right-color: #eb7e2d !important;border-right-style: solid !important;border-top-color: #eb7e2d !important;border-top-style: solid !important;border-bottom-color: #eb7e2d !important;border-bottom-style: solid !important;}&#8221;][vc_column][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243; css=&#8221;.vc_custom_1461851120811{border-right-width: 1px !important;border-right-color: #444444 !important;border-right-style: solid !important;}&#8221;][vc_column_text css=&#8221;.vc_custom_1485945692784{margin-top: 20px !important;}&#8221;]<\/p>\n<h2>NEBuilder Assembly Tool<\/h2>\n<p>Need help designing primers for DNA Assembly? Try <strong>NEBuilder Assembly Tool<\/strong>![\/vc_column_text][vc_column_text]<\/p>\n<p style=\"text-align: center;\"><span class=\"bildunterschrift\"><a href=\"http:\/\/nebuilder.neb.com\/\" target=\"_blank\" rel=\"noopener noreferrer\"><img decoding=\"async\" class=\"aligncenter size-full wp-image-2018\" src=\"http:\/\/www.neb-online.de\/wp-content\/uploads\/2015\/04\/NEBuilder150.png\" alt=\"NEBuilder\" width=\"63\" height=\"64\" \/><\/a>Please click the icon.<br \/>\n<\/span><\/p>\n<p>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text css=&#8221;.vc_custom_1462180839469{padding-top: 20px !important;}&#8221;]<\/p>\n<h2>NEBioCalculator<\/h2>\n<p>Use this tool for your scientific calculations and conversions for DNA and RNA[\/vc_column_text][vc_column_text]<\/p>\n<p style=\"text-align: center;\"><span class=\"bildunterschrift\"><a href=\"http:\/\/nebiocalculator.neb.com\/\" target=\"_blank\" rel=\"noopener noreferrer\"><img decoding=\"async\" class=\"aligncenter size-full wp-image-2017\" src=\"http:\/\/www.neb-online.de\/wp-content\/uploads\/2015\/04\/NEBioCalculator150.png\" alt=\"NEBioCalculator150\" width=\"62\" height=\"64\" \/><\/a>Please click the icon.<br \/>\n<\/span><\/p>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][\/vc_row][vc_row][vc_column][\/vc_column][vc_separator color=&#8221;black&#8221;][vc_column_text]<\/p>\n<h2>Comparison NEBuilder and Gibson Assembly<\/h2>\n<p>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;2\/3&#8243;][vc_column_text]<strong>NEBuilder HiFi DNA Assembly Master Mix offers improved efficiency and accuracy over Gibson Assembly with different numbers of fragments.<\/strong>[\/vc_column_text][vc_column_text]<span class=\"bildunterschrift\"><em>Reactions were set up in a 2- and 6- fragment assembly reaction according to recommended reaction conditions. <\/em><\/span>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_single_image image=&#8221;1536&#8243; img_size=&#8221;full&#8221; alignment=&#8221;center&#8221; img_link_large=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_separator][vc_row_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_single_image image=&#8221;1542&#8243; img_size=&#8221;full&#8221; alignment=&#8221;center&#8221; img_link_large=&#8221;yes&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;2\/3&#8243;][vc_column_text]<strong>NEBuilder HiFi DNA Assembly Master Mix offers improved efficiency and accuracy over Gibson Assembly, with lower amounts of DNA by increasing overlaps.<\/strong>[\/vc_column_text][vc_column_text]<span class=\"bildunterschrift\"><em>Reactions were set up in a 4- fragment assembly reaction according to recommended reaction conditions. Amount of DNA and size of overlap is shown. <\/em><\/span>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_separator][vc_row_inner][vc_column_inner width=&#8221;2\/3&#8243;][vc_column_text]<strong>NEBuilder HiFi DNA Master Mix offers improved fidelity over Gibson Assembly Master Mix<\/strong>[\/vc_column_text][vc_column_text]<span class=\"bildunterschrift\"><em>Fidelity of assembled products was compared between NEBuilder HiFi DNA Assembly Master Mix (NEB #E2621) and Gibson Assembly Master Mix (NEB #E2611). Experiments were performed using various fragment and vector sizes following suggested protocols. Experiments D and E did not yield any colonies with Gibson Assembly. Experiments H and J were performed with fragments containing 3\u00b4 restriction enzyme mismatches. Experiments B through G vary because sequences of fragments are different. <\/em>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_single_image image=&#8221;1546&#8243; img_size=&#8221;full&#8221; alignment=&#8221;center&#8221; img_link_large=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_separator color=&#8221;black&#8221;][vc_single_image image=&#8221;10701&#8243; img_size=&#8221;full&#8221; alignment=&#8221;center&#8221; onclick=&#8221;custom_link&#8221; img_link_target=&#8221;_blank&#8221; link=&#8221;https:\/\/www.neb.com\/forms\/nebuilder-hifi-dna-assembly-master-mix-sample-request-form&#8221;][vc_separator color=&#8221;black&#8221;][vc_row][\/vc_row][vc_column][\/vc_column][vc_column_text css=&#8221;.vc_custom_1461845865060{margin-top: 20px !important;padding-top: -20px !important;}&#8221;]<\/span><\/p>\n<h2>Videos<\/h2>\n<p>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_video link=&#8221;https:\/\/youtu.be\/LwOYvWTm7kE&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_video link=&#8221;https:\/\/youtu.be\/8_-t5xtJ3y8&#8243;][\/vc_column_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_video link=&#8221;https:\/\/youtu.be\/4wv4ETxL5PE&#8221;][\/vc_column_inner][\/vc_row_inner][vc_row_inner css=&#8221;.vc_custom_1461845782523{margin-top: -20px !important;}&#8221;][vc_column_inner width=&#8221;1\/3&#8243;][vc_column_text]<strong>Introduction to NEBuilder HiFi DNA Assembly<\/strong>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_column_text]<strong>Primer Design and Fragment Assembly Using NEBuilder HiFi DNA Assembly or Gibson Assembly<\/strong>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=&#8221;1\/3&#8243;][vc_column_text]<strong>NEBuilder HiFi DNA Assembly: Bridging dsDNA with a ssDNA Oligo<\/strong>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row][vc_column][vc_separator color=&#8221;black&#8221;][\/vc_column][\/vc_row][vc_row css=&#8221;.vc_custom_1461915371370{margin-right: -1px !important;margin-left: -1px !important;border-top-width: 2px !important;border-right-width: 2px !important;border-bottom-width: 2px !important;border-left-width: 2px !important;padding-top: 20px !important;padding-bottom: 20px !important;background-color: #f4f1e7 !important;border-left-color: #eb7e2d !important;border-left-style: solid !important;border-right-color: #eb7e2d !important;border-right-style: solid !important;border-top-color: #eb7e2d !important;border-top-style: solid !important;border-bottom-color: #eb7e2d !important;border-bottom-style: solid !important;}&#8221;][vc_column][vc_row_inner][vc_column_inner][vc_column_text css=&#8221;.vc_custom_1466499826080{padding-top: 10px !important;}&#8221;]<\/p>\n<h2>3 Application Notes<\/h2>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/6&#8243;][vc_single_image image=&#8221;8948&#8243; img_size=&#8221;full&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;5\/6&#8243;][vc_column_text]<strong>1.<\/strong> <a href=\"https:\/\/international.neb.com\/-\/media\/nebus\/files\/application-notes\/appnote_ssoligo_bridging_dsdna_crispr_sgrna.pdf?rev=e2a4ab38e14742229d1edcffc1b561f2\" target=\"_blank\" rel=\"noopener noreferrer\">Construction of an sgRNA-Cas9 expression vector via single-stranded DNA oligo bridging of double-stranded DNA fragments E5520<\/a><\/p>\n<p><strong>2.<\/strong> <a href=\"https:\/\/international.neb.com\/-\/media\/nebus\/files\/application-notes\/appnote_improved_methods_for_sdm_using_nebuilder_hifi_dna_assembly_master_mix.pdf?rev=7255867ee44d4936afb1cb9f87af9111\" target=\"_blank\" rel=\"noopener noreferrer\">Improved methods for site directed mutagenesis using NEBuilder HiFi DNA Assembly Master Mix<\/a><\/p>\n<p><strong>3.<\/strong> <a href=\"https:\/\/international.neb.com\/-\/media\/nebus\/files\/application-notes\/improved-method-for-assembly-of-linear-yeast-expression-e5520.pdf?rev=020d2d8a4d6d4d1b8238b2094e3ecb92&amp;hash=618DA6AB555803824BCE90B71D6E0287\" target=\"_blank\" rel=\"noopener noreferrer\">Improved method for assembly of linear yeast expression E5520<\/a>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][\/vc_row][vc_row][vc_column][vc_separator color=&#8221;black&#8221;][vc_column_text]<\/p>\n<h2>Available Products<\/h2>\n<p><strong>Order your favorite product for Cloning 2.0!<\/strong><\/p>\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=\"300\" 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=\"160\">SIZE<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" colspan=\"1\" width=\"200\">PRICE<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" colspan=\"1\" width=\"200\">MORE INFO<\/th>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\">NEBuilder HiFi DNA Assembly Cloning Kit<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E5520S<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">10 rxns<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"10px\"><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/E5520@\"><img decoding=\"async\" class=\"size-full wp-image-4375 aligncenter\" src=\"http:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/05\/shop_icon1.gif\" alt=\"shop_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"21px\"><a href=\"https:\/\/international.neb.com\/products\/E5520\" target=\"_blank\" rel=\"noopener noreferrer\"><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" src=\"https:\/\/www.neb-online.de\/enews\/enews_NEB_button_info.png\" alt=\"Information Produit\" width=\"20\" height=\"20\" \/><\/a><\/td>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\">NEBuilder HiFi DNA Assembly Master Mix<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E2621S\/L\/X<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">10\/50\/250 rxns<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"10px\"><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/e2621@\"><img decoding=\"async\" class=\"size-full wp-image-4375 aligncenter\" src=\"http:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/05\/shop_icon1.gif\" alt=\"shop_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"21px\"><a href=\"https:\/\/international.neb.com\/products\/E2621\" target=\"_blank\" rel=\"noopener noreferrer\"><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" src=\"https:\/\/www.neb-online.de\/enews\/enews_NEB_button_info.png\" alt=\"Information Produit\" width=\"20\" height=\"20\" \/><\/a><\/td>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\">NEBuilder HiFi DNA Assembly Bundle for Large Fragments<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E2623S<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">20 rxns<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"10px\"><a href=\"https:\/\/shop.neb-online.fr\/en\/art\/e2623@\"><img decoding=\"async\" class=\"size-full wp-image-4375 aligncenter\" src=\"http:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/05\/shop_icon1.gif\" alt=\"shop_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"21px\"><a href=\"https:\/\/international.neb.com\/products\/mE2623\" target=\"_blank\" rel=\"noopener noreferrer\"><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" src=\"https:\/\/www.neb-online.de\/enews\/enews_NEB_button_info.png\" alt=\"Information Produit\" width=\"20\" height=\"20\" \/><\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]<\/p>\n<p>[\/vc_column_text][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p>[vc_row][vc_column][vc_column_text]NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5\u00b4 and 3\u00b4 restriction enzyme mismatches. This flexible kit enables simple and fast Seamless Cloning utilizing a new proprietary high-fidelity polymerase. Find out why NEBuilder HiFi is the next generation of DNA assembly and cloning.[\/vc_column_text][vc_separator][vc_column_text] NEBuilder HiFi DNA Assembly Workflow [\/vc_column_text][vc_row_inner&#8230;  <a class=\"excerpt-read-more\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/nebuilder-hifi-dna-assembly\/\" title=\"Read NEBuilder HiFi DNA Assembly\">Read more &raquo;<\/a><\/p>\n","protected":false},"author":15,"featured_media":1984,"parent":2066,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"page-sidebar-cloning-synthetic-biology.php","meta":{"footnotes":""},"class_list":["post-8955","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>NEBuilder HiFi DNA 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\/nebuilder-hifi-dna-assembly\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"NEBuilder HiFi DNA Assembly - New England Biolabs France\" \/>\n<meta property=\"og:description\" content=\"[vc_row][vc_column][vc_column_text]NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5\u00b4 and 3\u00b4 restriction enzyme mismatches. This flexible kit enables simple and fast Seamless Cloning utilizing a new proprietary high-fidelity polymerase. Find out why NEBuilder HiFi is the next generation of DNA assembly and cloning.[\/vc_column_text][vc_separator][vc_column_text] NEBuilder HiFi DNA Assembly Workflow [\/vc_column_text][vc_row_inner... 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