Have a cloning project coming up and not sure which technique to use? Benchfly's Michael Gage recommends isothermal assembly reaction. While he was recently trying to attempt a five-point ligation, Gage says he decided to give the technique a shot. Basically, he says, the technique works by doing PCR using primers "such that each successive piece that you want to stitch together has about 20-30 bp of homology to the next piece." After the five-point ligation with isothermal assembly, about 50 percent of Gage's 100-200 colonies were correct. Isothermal assembly reaction is a "cloning hero," he says.
Need to Clone? Try This!
Oct 15, 2010