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From 2012.igem.org

Achievements and Failures

Successfully cloned and submitted 14 biobricks into the registry Successfully cloned over 32 more Biobricks waiting to be transferred into pSB1C3 plasmids Using PCR we obtained wbbC gene from BL21 genome Created GlycoBase; a database containing a list of over 100 different glycosyltransferase enzymes mainly from E.coli strains demonstrating the vast linkages we could achieve with just E.coli enzymes Created GlycoWeb, an interface for an online user to access Glycobase. Glycoweb has the ability to tell you which enzymes are needed to create your bespoke polysaccharide. Created GlycoApp; an application for a handheld device to access the Glycobase giving the user distance access and enhanced ordering capabilities Considered human practice approaches and discussed with several experts the human practice considerations, hosted a human practice panel and discussed with the wider community at Café Scientifique. Discussed our project with various businesses including Shell and we received letters of support from DSTL, ISCA Biochemical Ltd. Demonstrated that….. Successfully expressed this polysaccharide….. (to be cont….) Made promoter-RBS-gene-terminator constructs for two of the genes with more in construction We managed to obtain 7 of 9 fragments for Gibson assembly through PCR. The different annealing temperatures of the primers made this hard but showed how important the preparation for Gibson has to be In touching distance of completing our operons. Only time stopped this happening. We were one construct away from being able to complete our 3 gene inducible plasmid. Synthesis of a previously well characterised small polysaccharide, which can be physically and also biochemically characterised, to show that the product will act in biological systems as expected. Once a fully characterised polysaccharide is produced, we would be in a much stronger position to offer novel products and to start to address the exciting possibilities of our method.