Team:TU-Eindhoven/Protocols

From 2012.igem.org

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<h3>BioBrick<sup>TM</sup> construction</h3>
<h3>BioBrick<sup>TM</sup> construction</h3>
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<p>Many protcols for assembling BioBricks<sup>TM</sup> are already available, like Standard Assembly and 3A-assembly. These are suited for the assembly of standardized bricks, however, we created our BioBricks<sup>TM</sup> out of non-standard material and when working with yeast we used <span class="red">non-standard vectors</span> to carry the protein coding sequences. In other words, most of our <span class="red">cloning </span>steps had to be done <span class="red">the old-fashioned way</span> and consequently the yields were lower than with standard assembly. The solution was to use more DNA and longer incubation for digestion. This guaranteed a proper yield at the end of the process. The detail can be found in our <html><a href="https://static.igem.org/mediawiki/2012/d/dd/BioBrickprotocol.pdf">high-efficiency BioBrick<sup>TM</sup> assembly protocol</a></html>. This includes <span class="red">digestion, ligation and transformation</span>.</p>
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<p>Many protcols for assembling BioBricks<sup>TM</sup> are already available, like Standard Assembly and 3A-assembly. These are suited for the assembly of standardized bricks, however, we created our BioBricks<sup>TM</sup> out of non-standard material and when working with yeast we used <span class="red">non-standard vectors</span> to carry the protein coding sequences. In other words, most of our <span class="red">cloning </span>steps had to be done the <span class="red">old-fashioned way</span> and consequently the yields were lower than with standard assembly. The solution was to use more DNA and longer incubation for digestion. This guaranteed a proper yield at the end of the process. The detail can be found in our <html><a href="https://static.igem.org/mediawiki/2012/d/dd/BioBrickprotocol.pdf">high-efficiency BioBrick<sup>TM</sup> assembly protocol</a></html>. This includes <span class="red">digestion, ligation and transformation</span>.</p>

Revision as of 22:13, 26 September 2012