Team:Washington/Magnetosomes/Methods
From 2011.igem.org
Method of Extraction of Essential Magnetosome Genes
A genome prep was provided to our group by the Komeli lab at Berkeley and the sequence of the AMB-1 genome has been documented online [1]. This allowed us to quickly design pairs of primers that would extract the mamAB operon out in groups of 1-3 genes, trying to keep the chunks around 2kb long. These primers were ordered with an extra 20bp on their ends with homology to the bglbrk prefix on the forward primer and the bglbrk suffix on the reverse. This way, once the primers were used to extract the operon fragments, they could be cloned into a pGA vector via gibson assembly. Thus we broke up the operon into the groups shown below and submitted them as parts to the registry.