Solid-phase cloning for high-throughput assembly of single and multiple DNA parts

摘要: We describe solid-phase cloning (SPC) for high-throughput assembly of expression plasmids. Our method allows PCR products to be put directly into a liquid handler capture and purification using paramagnetic streptavidin beads conversion constructs by subsequent reactions. present robust automated protocol restriction enzyme based SPC its performance the >60 000 unique human gene fragments vectors. In addition, we report on SPC-based single-strand applications where exact control sequence between is needed or multiple inserts are assembled. this approach, solid support head-to-tail DNA hybridization polymerase fill-in. The usefulness was demonstrated >150 with up four parts at an average success rate above 80%. several suggest it particularly suitable efforts laboratory workstations.