Plant & Animal Genomes XIII Conference Plant & Animal Genomes XIII ConferenceJanuary 15-19, 2005
Town & Country Convention Center
San Diego, CA
David Mead , Melodee Patterson , Ron Godiska
Large sections of viral, microbial, and eukaryotic genomes are refractory to cloning with conventional host-vector systems. Most vectors induce transcription and translation of inserted fragments, leading to instability of certain classes of DNA sequences or gene products. This apparently “unclonable” DNA results in sequence “stacking”, clone gaps, or a complete inability to clone particular regions, including AT- or GC-rich sequences, toxic genes, nucleotide repeats, hairpins, and strong promoters. Lucigen’s CloneSmart® series of transcription-free vectors for plasmid, cosmid, fosmid, and BAC cloning greatly alleviates these problems. Use of these vectors dramatically improves insert stability and eliminates cloning bias in constructing genome libraries, thus reducing finishing costs in genome sequencing projects. Lucigen has also developed methods to clone genes and produce highly complex libraries from as little as a few nanograms of DNA. The latest Lucigen products, techniques, and technologies will be reviewed with specific examples in genomic cloning. A new procedure will also be discussed that increases recombinant yields up to 10-fold using any cloning vector compatible with E. coli competent cell strains.