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<h1>Purification</h1>
<h1>Purification</h1>
The usual procedure for purification of folded origamis uses agarose gel electrophoresis. Thereby the main product is separated from single staples and some misfolded structures, e.g. stabely linked dimers. While electrophoresis is a helpful tool for analysis of the folding products, for preparative means it has the disadvantages of low yields (estimated to be roughly 10% of the loaded structure) and, more importantly, the samples are contaminated with ethidium bromide. We investigated our origami with gel electrophoresis and found only one weak band in addition to the correctly folded structure. With such few misfolding events, the aim of purification was only to remove unused staples. This could be reached by using size exclusion filters with 100kDa molecular weight cutoff. This new procedure saves time (ca. 1/2 hour compared to several hours), is cheaper then resolubilization out of agarose gels, loss of product is significantly reduced and the origamis are not contaminated with intercalators. <br>
The usual procedure for purification of folded origamis uses agarose gel electrophoresis. Thereby the main product is separated from single staples and some misfolded structures, e.g. stably linked dimers. While electrophoresis is a helpful tool for analysis of the folding products, for preparative means it has the disadvantages of low yields (estimated to be roughly 10% of the loaded structure) and, more importantly, the samples are contaminated with ethidium bromide. We investigated our origami with gel electrophoresis and found only one weak band in addition to the correctly folded structure. With such few misfolding events, the aim of purification was only to remove unused staples. This could be reached by using size exclusion filters with 100kDa molecular weight cutoff. This new procedure saves time (ca. 1/2 hour as compared to several hours), is less expensive than resolubilization out of agarose gels, loss of product is significantly reduced, and the origamis are not contaminated with intercalators. <br>
For detailed information on the procedure, please read the [http://openwetware.org/wiki/Biomod/2011/TUM/TNT/LabbookA/Purification_of_origamis_by_size_exclusion_filtration#Purification_of_origamis_by_size_exclusion_filtration instruction] in our labbook. Results of the analysis of filtered structures with agarose gel electrophoresis can be found [http://openwetware.org/wiki/Biomod/2011/TUM/TNT/Results/Folding_Purification#Purification_with_size_excluscion_filters here].  
For detailed information on the procedure, please read the [http://openwetware.org/wiki/Biomod/2011/TUM/TNT/LabbookA/Purification_of_origamis_by_size_exclusion_filtration#Purification_of_origamis_by_size_exclusion_filtration instruction] in our labbook. Results of the analysis of filtered structures with agarose gel electrophoresis can be found [http://openwetware.org/wiki/Biomod/2011/TUM/TNT/Results/Folding_Purification#Purification_with_size_excluscion_filters here].  


Revision as of 10:26, 29 October 2011

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