Figure 1.

A streamlined BioBrick assembly protocol. The pairwise ligation of standardized parts was optimized for parallel processing on 96-well plates and thermocycler heat blocks. Plasmids with BioBrick-formatted parts are double-digested with restriction mix A or B, triple-ligated into a new vector backbone and transformed into competent cells. Transformants are screened by colony PCR and inoculated into deep-well plates for robotic or manual DNA miniprep. Plasmid DNA is diluted to a standard concentration and stored for further assembly rounds.