Application of Long-distance PCR to Restriction Site Mapping of a Cloned DNA Fragment on the λEMBL3 Phage Vector

Abstract

Long-distance PCR was applied to rapidly map the restriction sites of long inserts cloned on λEMBL3 phage vector. The restriction sites of 9 of 15 enzymes were completely assigned in a model experiment within 14 h, including 8 h for the PCR amplification. This method was found particularly useful for genomic DNA cloning when the partial sequence of the corresponding cDNA is known.