Registry of standard biological parts/PCR-based assembly


 * Automated PCR primer design for worm genes - This is a good example of a simple CAD tool to encourage assembly. You could imagine something similar to auto-generate the primers necessary for PCR-based assembly of standard parts.
 * Sauer:Stitching Genes by PCR
 * patent on some of this stuff

Demo
Going to get a better feel for this by doing a few biobrick constructions using PCR-assembly
 * Registry of standard biological parts/PCR-based assembly/Protocol


 * 1) J04500 + I13501 = J04450
 * 2) R0010 + I13504 = J04430


 * 1) F2620 + P1010 + E0240
 * 2) *will try this one as a 3-way

UDG
Can use UDG (NEB USER enzyme) to replace all restriction digests and could also eliminate secondary PCRs also. You would just need to design your oligos to contain a U in the right location, assuming that there'll be a T somewhere in the vacinity of the ends of the parts you are trying to combine.

In-Fusion

 * Clontech In-Fusion PCR cloning -- this could be used for the last step so that we don't need to do a digest/ligation at any point in the process.
 * Using In-fusion to combine multiple parts
 * Seamless cloning and gene fusion review Lu-2005
 * Useful forum discussion
 * Patents: 1, 2 (might be same as 1, title same though filing date is later), 3
 * This reference describes a system similar to In-Fusion using the vaccinia virus DNA polymerase Evans-2007