Ion pair-reversed phase HPLC approach facilitates subcloning of PCR products and screening of recombinant colonies

The isolation of a single DNA molecule for cloning is technically difficult , and the subsequent screening of colonies for recombinant DNA clones is ti me consuming. Ion pair-reversed phase HPLC (IP-RP-HPLC) analysis of nucleic acids improves the resolution and isolation of PCR products to be cloned W e demonstrate that PCR products analyzed and collected using the IP-RP-HPLC approach (WAVE(TM)DNA Fragment Analysis System) can be cloned directly int o a plasmid vector In addition, we demonstrate that when IP-RP-HPLC analysi s is extended to the colony screening process, the time required for these procedures is reduced.