{"id":1810,"date":"2021-06-05T16:46:20","date_gmt":"2021-06-05T16:46:20","guid":{"rendered":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/?page_id=1810"},"modified":"2021-08-02T12:52:02","modified_gmt":"2021-08-02T12:52:02","slug":"recombination-based-cloning-slice","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/services\/strategy-and-construct-design\/recombination-based-cloning-slice\/","title":{"rendered":"Recombination-based cloning: SLiCE"},"content":{"rendered":"\n<div class=\"vf-grid | vf-grid__col-3\"><div class=\"vf-grid__col--span-2\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p><strong>SLiCE<\/strong> cloning stands for <strong>S<\/strong>eamless <strong>L<\/strong>igation <strong>C<\/strong>loning <strong>E<\/strong>xtract and is based on an <em>in vitro<\/em> recombination process directly in <em>E. coli<\/em> cell extract. To enhance the cloning efficiency, this extract is usually prepared from an <em>E. coli<\/em> strain that expresses components of the \u03bb prophage Red\/ET recombination system. In a first step you\u2019ll perform a PCR with primers that contain 5\u2019 extensions to create a PCR fragment with short homologies to the area of the vector where you\u2019d like to insert your gene of interest. The vector can be linearized via PCR as well (or alternatively, with a restriction enzyme). Both vector and insert fragment are mixed with the cell extract to perform the SLiCE reaction, after which the mixture is transformed into an <em>E. coli<\/em> cloning strain.<\/p>\n\n<\/div>\n<\/div>\n\n\n<div><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<\/div>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":1,"featured_media":0,"parent":971,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"template-title-left-aligned.php","meta":{"_acf_changed":false,"footnotes":""},"embl_taxonomy":[],"class_list":["post-1810","page","type-page","status-publish","hentry"],"acf":[],"embl_taxonomy_terms":[],"_links":{"self":[{"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/pages\/1810","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/comments?post=1810"}],"version-history":[{"count":3,"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/pages\/1810\/revisions"}],"predecessor-version":[{"id":2178,"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/pages\/1810\/revisions\/2178"}],"up":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/pages\/971"}],"wp:attachment":[{"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/media?parent=1810"}],"wp:term":[{"taxonomy":"embl_taxonomy","embeddable":true,"href":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/wp-json\/wp\/v2\/embl_taxonomy?post=1810"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}