{"id":1824,"date":"2015-04-24T07:35:17","date_gmt":"2015-04-24T05:35:17","guid":{"rendered":"http:\/\/www.neb-online.fr\/?page_id=1824"},"modified":"2022-11-07T18:25:29","modified_gmt":"2022-11-07T17:25:29","slug":"cloning-synthetic-biology","status":"publish","type":"page","link":"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/","title":{"rendered":"Cloning \/ Synthetic Biology"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row][vc_column][vc_column_text]<\/p>\n<h2>Molecular Cloning<\/h2>\n<p>Molecular cloning refers to the process by which recombinant DNA molecules are produced and transformed into a host organism, where they are replicated.[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_single_image image=&#8221;1404&#8243; img_size=&#8221;full&#8221;]<a class=\"btn btn-standard form-btn\"  href=\"\/wp-content\/uploads\/2015\/04\/NEB_Klonierung_CloningTimeline.pdf\" style=\"margin-left:auto;margin-right:auto;\">Download image as PDF<\/a>[\/vc_column][\/vc_row][vc_row content_placement=&#8221;middle&#8221;][vc_column width=&#8221;1\/2&#8243;][vc_column_text]<\/p>\n<h3>How does it work?<\/h3>\n<p>A molecular cloning reaction is typically comprised of the following two components:<\/p>\n<ul>\n<li>The DNA fragment of interest to be replicated<\/li>\n<li>A vector\/plasmid backbone that contains all of the components for replication in the host<\/li>\n<\/ul>\n<p>DNA of interest, such as a gene, regulatory element(s), or operon, etc., is prepared for cloning by excising it out of the source DNA using <a title=\"Restriction Enzymes\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/\">restriction enzymes<\/a>, copying it using the <a title=\"PCR and DNA Amplification\" href=\"http:\/\/www.neb-online.fr\/en\/neb-en\/pcr-and-dna-amplification\/\">Polymerase Chain Reaction (PCR)<\/a>, or <a title=\"DNA Assembly\" href=\"http:\/\/www.neb-online.fr\/en\/neb-en\/cloning-synthetic-biology\/dna-assembly\/\">assembling<\/a> it from individual oligonucleotides. At the same time, a plasmid vector is prepared in linear form using restriction enzymes or PCR. The plasmid is a small, circular piece of DNA that is replicated within the host, and exists separately from the host\u2019s chromosomal or genomic DNA. By physically joining the DNA of interest to the plasmid vector through phosphodiester bonds, the DNA of interest becomes part of the new recombinant plasmid and is replicated by the host.[\/vc_column_text][\/vc_column][vc_column width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;1297&#8243; img_size=&#8221;full&#8221;]<a class=\"btn btn-standard form-btn\"  href=\"\/wp-content\/uploads\/2015\/04\/NEB_Klonierung_CloningWorkflows.pdf.png\" style=\"margin-left:auto;margin-right:auto;\">Download image as PDF<\/a>[\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]Plasmid vectors allow the DNA of interest to be copied in large amounts and, often, provide the necessary control elements to be used to direct transcription and translation of the cloned DNA. As such, they have become the workhorse for many molecular methods, such as <a title=\"Proteinexpression\" href=\"https:\/\/www.neb-online.fr\/en\/neb-en\/protein-expression\/\">protein expression<\/a>, <a title=\"Cellular Analysis (SNAP-Tag)\" href=\"http:\/\/www.neb-online.fr\/en\/neb-en\/cellular-analysis-snap-tag\/\">gene expression studies<\/a>, and functional analysis of biomolecules.<\/p>\n<p>During the cloning process, the ends of the DNA of interest and the vector have to be modified to make them compatible for joining through the action of a <a title=\"DNA Ligases\" href=\"https:\/\/www.neb-online.fr\/en\/t4-dna-ligase\/\">DNA ligase<\/a>, recombinase, or\u00a0<em>in vivo<\/em>\u00a0DNA repair mechanism. These steps typically utilize enzymes, such as <a title=\"DNA Modifying Enzymes\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-modifying-enzymes\/\">nucleases, phosphatases, kinases and\/or ligases<\/a>. Many cloning methodologies and, more recently, kits have been developed to simplify and standardize these processes.[\/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<h2>Synthetic Biology<\/h2>\n<p>Synthetic Biology is a more recent expansion of the biotechnology field, in which genes and proteins are viewed as parts or devices, with the goal of re-designing and\/or assembling these parts in novel ways to create a new and useful functionality. Recent advances in biofuels generation, production of biochemicals, and understanding the minimal genome all benefit from synthetic biological approaches.<\/p>\n<p>Often these projects rely on the ordered <a title=\"DNA Assembly\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/\">assembly<\/a> of multiple DNA sequences to create large, artificial DNA structures. To this end, methods have evolved to simplify this process.<\/p>\n<p><a title=\"DNA Assembly\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/dna-assembly\/nebuilder-hifi-dna-assembly\/\">NEBuilder\u00ae HiFi DNA Assembly<\/a> and Gibson Assembly\u00ae can be used to create many functional DNA structures, from a simple joining of two metabolic genes, all the way up to the creation of an artificial genome.[\/vc_column_text][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p>[vc_row][vc_column][vc_column_text] Molecular Cloning Molecular cloning refers to the process by which recombinant DNA molecules are produced and transformed into a host organism, where they are replicated.[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_single_image image=&#8221;1404&#8243; img_size=&#8221;full&#8221;][\/vc_column][\/vc_row][vc_row content_placement=&#8221;middle&#8221;][vc_column width=&#8221;1\/2&#8243;][vc_column_text] How does it work? A molecular cloning reaction is typically comprised of the following two components: The DNA fragment of interest to be replicated A&#8230;  <a class=\"excerpt-read-more\" href=\"https:\/\/www.neb-online.fr\/en\/cloning-synthetic-biology\/\" title=\"Read Cloning \/ Synthetic Biology\">Read more &raquo;<\/a><\/p>\n","protected":false},"author":15,"featured_media":1849,"parent":0,"menu_order":0,"comment_status":"open","ping_status":"closed","template":"page-sidebar-cloning-synthetic-biology.php","meta":{"footnotes":""},"class_list":["post-1824","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>Cloning \/ Synthetic Biology - 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\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Cloning \/ Synthetic Biology - New England Biolabs France\" \/>\n<meta property=\"og:description\" content=\"[vc_row][vc_column][vc_column_text] Molecular Cloning Molecular cloning refers to the process by which recombinant DNA molecules are produced and transformed into a host organism, where they are replicated.[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_single_image image=&#8221;1404&#8243; img_size=&#8221;full&#8221;][\/vc_column][\/vc_row][vc_row content_placement=&#8221;middle&#8221;][vc_column width=&#8221;1\/2&#8243;][vc_column_text] How does it work? A molecular cloning reaction is typically comprised of the following two components: The DNA fragment of interest to be replicated A... 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