In order to build the large assembled structures using the designed nanoBricks, an assembler-type device is required. Such device allows the automation of frequent and repetitive liquid dispensing steps (pipetting) and thus forms an acceptable bridge between the digital version of the sought model and the real-life structure. It has been suggested by a number of colleagues that
HPLC purification of the linkers is necessary for their increased purity and efficiency in linking two nanoBricks together. Given the relatively high cost (
~$50, dependent on oligo length) of the resulting linker, a shift from global to local address space is desirable.
Global vs Local Address Space
Often in Biology, a global address space is used when conceiving DNA assembly strategies. This enables "one-pot" assemblies where all the components can be combined at once to form the product. As you can easily imagine, this requires strong binding affinities to be used and is not particularly robust to interfering molecules. In this particular context, a global address space signifies that each linker between nanoBricks must be unique an separate from one another in order to allow one-pot assembly. This significantly increases the cost of assembly of the large structure.
In order to keep the costs lower,
solid phase synthesis can be used to effectively implement a local address space in the assembly process. Using a Streptavidin-Biotin bond to anchor the first nanoBrick to the construction chamber (PDMS or Glass), the structure is seeded and then extended by flowing successive linkers and nanoBricks.