Team:Penn State/MSC Design
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Revision as of 23:42, 3 October 2012
Multiple Start Codons
A frustrating yet commonly observed phenomenon in the lab is the production of unexpected proteins. These occurrences may be explainable by multiple start codons in the mRNA strand. Codon slippage is a theory practically untouched by research, and this project aspires to shed some light on the issue.
Multiple Start Codons
Initial Construct
The initial construct for this project was designed using a basic vector and a relatively new product available from IDT (Integrated DNA Technologies), called gBlocks. gBlocks are synthesized dsDNA oligos with adjacent 40 base-pair overlap sequences. These oligos are particularly useful in multi-part constructions within a CBAR or Gibson Assembly reaction protocol.
Assembly
The oligos were ordered based on a digital representation of the desired plasmid sequence. The DNA sequence was manipulated through the free software ApE (A plasmid Editor) available through the University of Utah
Initial construction proved difficult. Whether this was due to inexperience or poor design, we cannot be sure. The final construct displaying a correct sequence has been assembled, however, and is giving us new insight into the codon slippage pheomenon.