{"id":2077,"date":"2015-04-27T10:59:23","date_gmt":"2015-04-27T10:59:23","guid":{"rendered":"http:\/\/www.neb-online.fr\/?page_id=2077"},"modified":"2022-11-10T16:33:29","modified_gmt":"2022-11-10T15:33:29","slug":"pcr-cloning-kit","status":"publish","type":"page","link":"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/pcr-cloning-kit\/","title":{"rendered":"PCR Cloning Kit"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row][vc_column][vc_column_text][\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column width=&#8221;1\/2&#8243; css=&#8221;.vc_custom_1428969316664{margin-bottom: 30px !important;}&#8221;][vc_column_text]The <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/e1202@\" target=\"_blank\" rel=\"noopener noreferrer\">NEB PCR Cloning Kit<\/a> contains an optimized 2X Cloning Master Mix with a proprietary ligation enhancer and a linearized vector that uses a novel mechanism for background colony suppression to give a low background.<\/p>\n<p>It allows simple and quick cloning of any PCR amplicon, whether the amplification reactions are performed with proofreading <a title=\"PCR and DNA Amplification\" href=\"http:\/\/www.neb-online.fr\/en\/neb-en\/pcr-and-dna-amplification\/\">DNA polymerases<\/a>, such as <a title=\"Q5 High Fidelity DNA Polymerase\" href=\"https:\/\/www.neb-online.fr\/en\/pcr-and-dna-amplification\/high-fidelity-pcr\/q5-high-fidelity-dna-polymerase\/\">Q5\u00ae<\/a> or <a title=\"Phusion DNA Polymerase\" href=\"https:\/\/www.neb-online.fr\/en\/pcr-and-dna-amplification\/high-fidelity-pcr\/phusion-dna-polymerase\/\">Phusion\u00ae<\/a> which produce blunt ends; or nonproofreading DNA polymerases, such as\u00a0<a title=\"Taq DNA Polymerases\" href=\"https:\/\/www.neb-online.fr\/en\/pcr-and-dna-amplification\/routine-pcr-en\/taq-dna-polymerases\/\"><em>Taq<\/em><\/a>\u00a0or\u00a0<em>Taq<\/em>\u00a0mixes (<a title=\"OneTaq DNA Polymerase\" href=\"https:\/\/www.neb-online.fr\/en\/pcr-and-dna-amplification\/routine-pcr-en\/onetaq-dna-polymerase\/\">One<em>Taq<\/em>\u00ae<\/a>, <a href=\"https:\/\/shop.neb-online.fr\/en\/art\/M0323@\" target=\"_blank\" rel=\"noopener noreferrer\">LongAmp\u2122\u00a0<em>Taq<\/em><\/a>) which produce single base overhangs. This is possible due to \u201cinvisible\u201d end polishing components in the master mix that are active during the ligation step.<\/p>\n<p>The kit also allows direct cloning from amplification reactions without purification, and works well whether or not the primers used in the PCR possess 5\u00b4-phosphate groups.[\/vc_column_text][\/vc_column][vc_column width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;1006&#8243; img_size=&#8221;full&#8221;][vc_column_text]<em>A 500 bp PCR product incubated with the linearized vector in a 3:1 ratio\u00a0according to recommended protocol. 2 \u03bcl of reaction was transformed into\u00a0provided NEB 10-beta Competent E. coli and 1\/20th of the outgrowth was\u00a0plated. Left plate serves as the control, with vector backbone only, right plate\u00a0contains PCR insert.<\/em>[\/vc_column_text][\/vc_column][vc_column css=&#8221;.vc_custom_1428969068440{margin-top: 20px !important;}&#8221;][vc_separator color=&#8221;custom&#8221; accent_color=&#8221;#919191&#8243;][\/vc_column][vc_column css=&#8221;.vc_custom_1428969002458{margin-top: 30px !important;}&#8221;][vc_column_text]<strong>Advantages<\/strong><\/p>\n<ul>\n<li>Easy cloning of all PCR products, including blunt and TA ends<\/li>\n<li>Fast cloning experiments with 5-minute ligation step<\/li>\n<li>Simplified screening with low\/no colony background and no blue\/white selection required<\/li>\n<li>Save time by eliminating purification steps<\/li>\n<li>Flanking restriction sites available for easy subcloning, including choice of two single digest options<\/li>\n<li>Provided analysis primers allow for downstream colony PCR screening or sequencing<\/li>\n<li>Ready-to-use kit components include 1 kb control amplicon, linearized cloning vector and single-use competent\u00a0<em>E. coli<\/em><\/li>\n<li>Longer shelf life (12 months), as compared to some commercially available products<\/li>\n<\/ul>\n<p>[\/vc_column_text][vc_separator color=&#8221;custom&#8221; accent_color=&#8221;#919191&#8243;][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text][\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]<span class=\"bildunterschrift\"><img fetchpriority=\"high\" decoding=\"async\" class=\"alignnone wp-image-9016 size-full\" src=\"http:\/\/www.neb-online.fr\/wp-content\/uploads\/E1202c-NEW.jpg\" alt=\"NEB PCR Cloning Kit pMIniT 2.0\" width=\"575\" height=\"884\" srcset=\"https:\/\/www.neb-online.fr\/wp-content\/uploads\/E1202c-NEW.jpg 575w, https:\/\/www.neb-online.fr\/wp-content\/uploads\/E1202c-NEW-195x300.jpg 195w\" sizes=\"(max-width: 575px) 100vw, 575px\" \/><br \/>\n<em><strong>pMiniT Vector Map<br \/>\n<\/strong><\/em><em>Map shown above displays the construct formed if no insert is present. Unique restriction sites are shown<\/em> <em>in black. Additional restriction sites that can be used for subcloning are shown in red. Expanded box below<\/em> <em>shows location of sequencing primers, restriction sites for subcloning, and placement of insertion site<\/em> <em>within the toxic minigene.<\/em><\/span>[\/vc_column_text][vc_separator color=&#8221;black&#8221;][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]<\/p>\n<h2>Available Products<\/h2>\n<table style=\"width: 793px; height: 174px;\" border=\"0\" cellpadding=\"5\">\n<tbody>\n<tr style=\"background-color: #f3b281;\">\n<th style=\"border-bottom: solid #eb7e2d;\" width=\"351\" height=\"40\">PRODUCT<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" width=\"73\">REF.<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" width=\"111\">SIZE<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" colspan=\"1\">PRICE<\/th>\n<th style=\"border-bottom: solid #eb7e2d;\" colspan=\"1\">MORE INFO<\/th>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\">NEB PCR Cloning Kit (incl. <em>E. coli<\/em> competent Cells)<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E1202 S<\/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\/e1202s\"><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<td style=\"border-bottom: 1px solid #eb7e2d;\" width=\"25\"><a href=\"https:\/\/international.neb.com\/products\/e1202-neb-pcr-cloning-kit\"><img decoding=\"async\" class=\"aligncenter\" src=\"http:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/04\/NEB_Info_icon.png\" alt=\"NEB Info_icon\" width=\"25\" height=\"25\" \/><\/a><\/td>\n<\/tr>\n<tr>\n<td style=\"border-bottom: 1px solid #eb7e2d;\">NEB PCR Cloning Kit (Without Competent Cells)<\/td>\n<td style=\"border-bottom: 1px solid #eb7e2d; text-align: center;\">E1203 S<\/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\/e1203s\"><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter wp-image-4375 size-full\" 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=\"25\"><a href=\"http:\/\/international.neb.com\/products\/e1203-neb-pcr-cloning-kit-without-competent-cells\"><img decoding=\"async\" class=\"aligncenter\" src=\"http:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/04\/NEB_Info_icon.png\" alt=\"NEB Info_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][vc_column_text][\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column width=&#8221;1\/2&#8243; css=&#8221;.vc_custom_1428969316664{margin-bottom: 30px !important;}&#8221;][vc_column_text]The NEB PCR Cloning Kit contains an optimized 2X Cloning Master Mix with a proprietary ligation enhancer and a linearized vector that uses a novel mechanism for background colony suppression to give a low background. It allows simple and quick cloning of any PCR amplicon, whether the amplification reactions are performed&#8230;  <a class=\"excerpt-read-more\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/pcr-cloning-kit\/\" title=\"Read PCR Cloning Kit\">Read more &raquo;<\/a><\/p>\n","protected":false},"author":15,"featured_media":1849,"parent":1824,"menu_order":0,"comment_status":"open","ping_status":"closed","template":"page-sidebar-cloning-synthetic-biology.php","meta":{"footnotes":""},"class_list":["post-2077","page","type-page","status-publish","has-post-thumbnail","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.5 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>PCR Cloning Kit - 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\/pcr-cloning-kit\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"PCR Cloning Kit - New England Biolabs France\" \/>\n<meta property=\"og:description\" content=\"[vc_row][vc_column][vc_column_text][\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column width=&#8221;1\/2&#8243; css=&#8221;.vc_custom_1428969316664{margin-bottom: 30px !important;}&#8221;][vc_column_text]The NEB PCR Cloning Kit contains an optimized 2X Cloning Master Mix with a proprietary ligation enhancer and a linearized vector that uses a novel mechanism for background colony suppression to give a low background. It allows simple and quick cloning of any PCR amplicon, whether the amplification reactions are performed... Read more &raquo;\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/pcr-cloning-kit\/\" \/>\n<meta property=\"og:site_name\" content=\"New England Biolabs France\" \/>\n<meta property=\"article:modified_time\" content=\"2022-11-10T15:33:29+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/www.neb-online.fr\/wp-content\/uploads\/2015\/04\/NEB_Klonierung_Header_klonierung_760px.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"760\" \/>\n\t<meta property=\"og:image:height\" content=\"200\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"twitter:label1\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data1\" content=\"3 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/\",\"url\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/\",\"name\":\"PCR Cloning Kit - New England Biolabs France\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/#website\"},\"primaryImageOfPage\":{\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/#primaryimage\"},\"image\":{\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/#primaryimage\"},\"thumbnailUrl\":\"https:\\\/\\\/www.neb-online.fr\\\/wp-content\\\/uploads\\\/2015\\\/04\\\/NEB_Klonierung_Header_klonierung_760px.jpg\",\"datePublished\":\"2015-04-27T10:59:23+00:00\",\"dateModified\":\"2022-11-10T15:33:29+00:00\",\"breadcrumb\":{\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/#breadcrumb\"},\"inLanguage\":\"en-US\",\"potentialAction\":[{\"@type\":\"ReadAction\",\"target\":[\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/\"]}]},{\"@type\":\"ImageObject\",\"inLanguage\":\"en-US\",\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/#primaryimage\",\"url\":\"https:\\\/\\\/www.neb-online.fr\\\/wp-content\\\/uploads\\\/2015\\\/04\\\/NEB_Klonierung_Header_klonierung_760px.jpg\",\"contentUrl\":\"https:\\\/\\\/www.neb-online.fr\\\/wp-content\\\/uploads\\\/2015\\\/04\\\/NEB_Klonierung_Header_klonierung_760px.jpg\",\"width\":760,\"height\":200,\"caption\":\"Klonierung Header\"},{\"@type\":\"BreadcrumbList\",\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/pcr-cloning-kit\\\/#breadcrumb\",\"itemListElement\":[{\"@type\":\"ListItem\",\"position\":1,\"name\":\"Cloning \\\/ Synthetic Biology\",\"item\":\"https:\\\/\\\/www.neb-online.fr\\\/en\\\/cloning-synthetic-biology\\\/\"},{\"@type\":\"ListItem\",\"position\":2,\"name\":\"PCR Cloning Kit\"}]},{\"@type\":\"WebSite\",\"@id\":\"https:\\\/\\\/www.neb-online.fr\\\/#website\",\"url\":\"https:\\\/\\\/www.neb-online.fr\\\/\",\"name\":\"New England Biolabs France\",\"description\":\"be INSPIRED, drive DISCOVERY, stay GENUINE\",\"potentialAction\":[{\"@type\":\"SearchAction\",\"target\":{\"@type\":\"EntryPoint\",\"urlTemplate\":\"https:\\\/\\\/www.neb-online.fr\\\/?s={search_term_string}\"},\"query-input\":{\"@type\":\"PropertyValueSpecification\",\"valueRequired\":true,\"valueName\":\"search_term_string\"}}],\"inLanguage\":\"en-US\"}]}<\/script>\n<!-- \/ Yoast SEO plugin. -->","yoast_head_json":{"title":"PCR Cloning Kit - New England Biolabs France","robots":{"index":"index","follow":"follow","max-snippet":"max-snippet:-1","max-image-preview":"max-image-preview:large","max-video-preview":"max-video-preview:-1"},"canonical":"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/pcr-cloning-kit\/","og_locale":"en_US","og_type":"article","og_title":"PCR Cloning Kit - New England Biolabs France","og_description":"[vc_row][vc_column][vc_column_text][\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column width=&#8221;1\/2&#8243; css=&#8221;.vc_custom_1428969316664{margin-bottom: 30px !important;}&#8221;][vc_column_text]The NEB PCR Cloning Kit contains an optimized 2X Cloning Master Mix with a proprietary ligation enhancer and a linearized vector that uses a novel mechanism for background colony suppression to give a low background. 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