{"id":354,"date":"2023-10-16T11:25:24","date_gmt":"2023-10-16T10:25:24","guid":{"rendered":"https:\/\/sites.dundee.ac.uk\/david-horn\/?page_id=354"},"modified":"2026-02-13T15:58:54","modified_gmt":"2026-02-13T15:58:54","slug":"stem-loop-rnai-cassettes","status":"publish","type":"page","link":"https:\/\/sites.dundee.ac.uk\/david-horn\/resources\/stem-loop-rnai-cassettes\/","title":{"rendered":"Stem loop RNAi cassettes"},"content":{"rendered":"\n<p>Cassette sequenced in full, February 2008<\/p>\n\n\n\n<p>Two versions are available, containing different multiple cloning sites separated by a 468 bp stuffer (pBluescript 117-585, including 125 bp of the \u03b2-galactosidase \u03b1-fragment)<\/p>\n\n\n\n<p>All of the restriction sites listed are unique.<\/p>\n\n\n\n<pre class=\"wp-block-preformatted\"><strong>MCS<sup>1\/2<\/sup><\/strong>\n\n<em>Kpn<\/em>I <em>Sma<\/em>I <em>Afl<\/em>II <em>Bam<\/em>HI\n  1  <em>GGTA<strong>CC<\/strong><\/em><strong>CGGG<\/strong> <em>CTTAA<strong>G<\/strong><\/em><strong>GATC C<\/strong>GAGATAGGG TTGAGTGTTG TTCCAGTTTG\n 51  GAACAAGAGT CCACTATTAA AGAACGTGGA CTCCAACGTC AAAGGGCGAA\n101  AAACCGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC ACCCTAATCA\n151  AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA ACCCTAAAGG\n201  GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA\n251  AGGAAGGGAA GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA\n301  GCGGTCACGC TGCGCGTAAC CACCACACCC GCCGCGCTTA ATGCGCCGCT\n351  ACAGGGCGCG TCCATTCGCC ATTCAGGCTG CGCAACTGTT GGGAAGGGCG\n401  ATCGGTGCGG GCCTCTTCGC TATTACGCCA GCTGGCGAAA GGGGGATGTG\n451  CTGCAAGGCG ATTAAGTTGG GTAACGCCAG GGTTT<strong>TCTAG <em>A<\/em><\/strong><em>AGCTT<\/em><strong>CTCG<\/strong>\n501  <strong>A<em>G<\/em><\/strong><em>GGCCC<\/em> \n\n<em>Xba<\/em>I <em>Hin<\/em>dIII <em>Xho<\/em>I <em>Apa<\/em>I\n<strong>MCS<sup>3\/4<\/sup><\/strong>\n<em>Kpn<\/em>I <em>Age<\/em>I <em>Cla<\/em>I <em>Mfe<\/em>I <em>Sma<\/em>I\nNote: <em>Age<\/em>I &amp; <em>Xma<\/em>I (isoschizomer of <em>Sma<\/em>I) give compatible overhangs; <em>Cla<\/em>I site blocked by Dam methylation\n\n  1  <em>GGT<strong>ACC<\/strong><\/em><strong>GGT<\/strong><em>A TCGAT<\/em><strong>CAATT G<\/strong><em>CCCGGG<\/em>TCC GAGATAGGGT TGAGTGTTGT\n 51  TCCAGTTTGG AACAAGAGTC CACTATTAAA GAACGTGGAC TCCAACGTCA\n101  AAGGGCGAAA AACCGTCTAT CAGGGCGATG GCCCACTACG TGAACCATCA\n151  CCCTAATCAA GTTTTTTGGG GTCGAGGTGC CGTAAAGCAC TAAATCGGAA\n201  CCCTAAAGGG AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG CCGGCGAACG\n251  TGGCGAGAAA GGAAGGGAAG AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG\n301  GCAAGTGTAG CGGTCACGCT GCGCGTAACC ACCACACCCG CCGCGCTTAA\n351  TGCGCCGCTA CAGGGCGCGT CCATTCGCCA TTCAGGCTGC GCAACTGTTG\n401  GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT ATTACGCCAG CTGGCGAAAG\n451  GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG GTTTTC<strong>AGAT<\/strong>\n501  <strong><em>CT<\/em><\/strong><em>TAA<strong>G<\/strong><\/em><strong>GCCG GC<em>C<\/em><\/strong><em>TCGAG<\/em><strong>TC TAGA<\/strong><em>GGGCCC<\/em>\n\n<em>Bgl<\/em>II <em>Afl<\/em>II <em>Fse<\/em>I <em>Xho<\/em>I <em>Xba<\/em>I <em>Apa<\/em>I<\/pre>\n","protected":false},"excerpt":{"rendered":"<p>Cassette sequenced in full, February 2008 Two versions are available, containing different multiple cloning sites separated by a 468 bp stuffer (pBluescript 117-585, including 125 bp of the \u03b2-galactosidase \u03b1-fragment) All of the restriction sites listed are unique. MCS1\/2 KpnI SmaI AflII BamHI 1 GGTACCCGGG CTTAAGGATC CGAGATAGGG TTGAGTGTTG TTCCAGTTTG 51 GAACAAGAGT CCACTATTAA AGAACGTGGA CTCCAACGTC AAAGGGCGAA [&hellip;]<\/p>\n","protected":false},"author":6,"featured_media":0,"parent":202,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"page-sub-landing.php","meta":{"_acf_changed":false,"footnotes":""},"class_list":["post-354","page","type-page","status-publish","hentry"],"blocksy_meta":[],"acf":[],"_links":{"self":[{"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/pages\/354","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/users\/6"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/comments?post=354"}],"version-history":[{"count":2,"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/pages\/354\/revisions"}],"predecessor-version":[{"id":586,"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/pages\/354\/revisions\/586"}],"up":[{"embeddable":true,"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/pages\/202"}],"wp:attachment":[{"href":"https:\/\/sites.dundee.ac.uk\/david-horn\/wp-json\/wp\/v2\/media?parent=354"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}