Ion pair-reversed phase HPLC approach facilitates subcloning of PCR products and screening of recombinant colonies
Shaw-Bruha, C.M.; Lamb, K.A.
Biotechniques 28(4): 794-797
2000
ISSN/ISBN: 0736-6205 PMID: 10769760 Document Number: 513631
The isolation of a single DNA molecule for cloning is technically difficult, and the subsequent screening of colonies for recombinant DNA clones is time consuming. Ion pair-reversed phase HPLC (IP-RP-HPLC) analysis of nucleic acids improves the resolution and isolation of PCR products to be cloned. We demonstrate that PCR products analyzed and collected using the IP-RP-HPLC approach (WAVETM DNA Fragment Analysis System) can be cloned directly into a plasmid vector. In addition, we demonstrate that when IP-RP-HPLC analysis is extended to the colony screening process, the time required for these procedures is reduced.