title: Short homologies efficiently generate detectable homologous recombination events.
abstract: When recombineering bacterial artificial chromosomes (BACs), it is common practice to design the ends of the donor molecule with 50 bp of homology specifying its insertion site. We demonstrate that desired recombinants can be produced using intermolecular homologies as short as 15 bp. Although the use of shorter donor end regions decreases total recombinants by several fold, the frequency of recombinants with correctly inserted donor molecules was high enough for easy detection by simple polymerase chain reaction (PCR) screening. This observation may have important implications for the design of oligonucleotides for recombineering, including significant cost savings, especially for high-throughput projects that use large quantities of primers.
authors.affiliation: School of Science, Monash University Malaysia, 46150 Bandar Sunway, Selangor, Malaysia; Department of Biotechnology, Malaysia University of Science and Technology, 47301 Petaling Jaya, Selangor Darul Ehsan, Malaysia.
authors.forename: Andrew N
authors.initials: AN
authors.lastname: Osahor
authors.affiliation: School of Science, Monash University Malaysia, 46150 Bandar Sunway, Selangor, Malaysia.
authors.forename: Chau-Yan
authors.initials: CY
authors.lastname: Tan
authors.affiliation: Department of Molecular Biology, Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia.
authors.forename: Edmund Ui-Hang
authors.initials: EU
authors.lastname: Sim
authors.affiliation: Laboratory of Microbial Ecology, Institute of Biological Sciences, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Choon-Weng
authors.initials: CW
authors.lastname: Lee
authors.affiliation: School of Science, Monash University Malaysia, 46150 Bandar Sunway, Selangor, Malaysia; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: kumaran.narayanan@monash.edu.
authors.forename: Kumaran
authors.initials: K
authors.lastname: Narayanan
mesh_terms: D001483:Base Sequence; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D059765:Homologous Recombination; D010957:Plasmids / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D016615:Telomere / Q000235:genetics
keywords: BACs; Chromosome; E. coli; Homology; Linear; Recombineering