A vector for purification-free cloning of polymerase chain reaction products

Conventional cloning requires the purification of restriction-enzyme-digested vectors prior to the ligation reaction. The purification often involves the separation of restriction fragments via electrophoresis, the cutting out of a piece of gel, and the gel extraction of the linearized vector. In addition to the loss of significant amounts of DNA, reduced cloning efficiency, time, and cost, these steps are also mutagenic to DNA and hazardous to humans. We developed a purification-free cloning vector pGT3 with a bright green fluorescent protein indicator that is suitable for TA cloning of polymerase chain reaction (PCR) products. PCR products were cloned into pGT3 efficiently without the gel purification steps.