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Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
Published on: January 8, 2015
New vectors for direct cloning of PCR products
J Cha1, W Bishai, S Chandrasegaran
1Department of Environmental Health Sciences, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, MD 21205.
Abstract:
We describe the construction of two new vectors for direct cloning of polymerase chain reaction (PCR) products. This was done by inserting a synthetic DNA fragment containing two adjacent XcmI sites between the Asp718 and BamHI sites of the M13mp18 and M13mp19 phages. Cleavage of these M13 derivatives with XcmI will result in a linearized vector with a single thymidine nucleotide at the 3' ends. Thus, these vectors would be very useful for direct cloning of PCR-generated products with high efficiency.
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