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Team:TU Darmstadt/Notebook/Labjournal/K1497013
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| - | <!--TYPO3SEARCH_begin--><div id="c137" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">K1497013 - Dihydroflavonol 4-reductase with strong RBS (B0034)</h1></div><div class="csc-textpic-text"><p>After we created BBa_K1497010, a ribosom binding site had to be added to come a step further for creating a functioning expression system. For this purpose, we used BBa_K1497010 as a template for PCR with RBS primers (Prefix_RBS_DFR-primer and DFR_Suffix-primer). The PCR product was then restricted with the restriction enzymes EcoRI and PstI and ligated into an equaly restricted pSB1C3 vector to create the biobrick BBa_K1497013. </p></div></div><div id="c138" class="csc-default"><div class="csc-textpic csc-textpic-intext-right"><div class="csc-textpic-imagewrap" data-csc-images="1" data-csc-cols="2"><figure class="csc-textpic-image csc-textpic-last"><img src="https://static.igem.org/mediawiki/parts/6/65/070814_RestriktionPlasmidprep060814_1.png" width="300" height="225" alt=""></figure></div><div class="csc-textpic-text"><p>Figure | + | <!--TYPO3SEARCH_begin--><div id="c137" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">K1497013 - Dihydroflavonol 4-reductase with strong RBS (B0034)</h1></div><div class="csc-textpic-text"><p>After we created BBa_K1497010, a ribosom binding site had to be added to come a step further for creating a functioning expression system. For this purpose, we used BBa_K1497010 as a template for PCR with RBS primers (Prefix_RBS_DFR-primer and DFR_Suffix-primer). The PCR product was then restricted with the restriction enzymes EcoRI and PstI and ligated into an equaly restricted pSB1C3 vector to create the biobrick BBa_K1497013. </p></div></div><div id="c138" class="csc-default"><div class="csc-textpic csc-textpic-intext-right"><div class="csc-textpic-imagewrap" data-csc-images="1" data-csc-cols="2"><figure class="csc-textpic-image csc-textpic-last"><img src="https://static.igem.org/mediawiki/parts/6/65/070814_RestriktionPlasmidprep060814_1.png" width="300" height="225" alt=""></figure></div><div class="csc-textpic-text"><p>Figure 1: gel electrophoresis of DFR in pSB1C3 digested and undigested: 1st lane contains undigested RBS_DFR in pSB1C3 (green box), 2nd lane contains RBS_DFR digested with Pst1 and EcoR1 (red box)</p></div></div></div><!--TYPO3SEARCH_end--> |
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Revision as of 23:36, 15 October 2014
K1497013 - Dihydroflavonol 4-reductase with strong RBS (B0034)
After we created BBa_K1497010, a ribosom binding site had to be added to come a step further for creating a functioning expression system. For this purpose, we used BBa_K1497010 as a template for PCR with RBS primers (Prefix_RBS_DFR-primer and DFR_Suffix-primer). The PCR product was then restricted with the restriction enzymes EcoRI and PstI and ligated into an equaly restricted pSB1C3 vector to create the biobrick BBa_K1497013.
Figure 1: gel electrophoresis of DFR in pSB1C3 digested and undigested: 1st lane contains undigested RBS_DFR in pSB1C3 (green box), 2nd lane contains RBS_DFR digested with Pst1 and EcoR1 (red box)
