title: Optical mapping of DNA: single-molecule-based methods for mapping genomes.
abstract: The technologies associated with DNA sequencing are rapidly evolving. Indeed, single-molecule DNA sequencing strategies are cheaper and faster than ever before. Despite this progress, every sequencing platform to date relies on reading the genome in small, abstract fragments, typically of less than 1000 bases in length. The overarching aim of the optical map is to complement the information derived from DNA sequencing by providing long-range context on which these short sequence reads can be built. This is typically done using an enzyme to target and modify at short DNA sequences of, say, six bases in length throughout the genome. By accurately placing these short pieces of sequence on long genomic DNA fragments, up to several millions of bases in length, a scaffold for sequence assembly can be obtained. This review focuses on three enzymatic approaches to optical mapping. Optical mapping was first developed using restriction enzymes to sequence-specifically cleave DNA that is immobilized on a surface. More recently, nicking enzymes have found application in the sequence-specific fluorescent labeling of DNA for optical mapping. Such covalent modification allows the DNA to be imaged in solution, and this, in combination with developing nanofluidic technologies, is enabling new high-throughput approaches to mapping. And, finally, this review will discuss the recent development of mapping with subdiffraction-limit precision using methyltransferase enzymes to label the DNA with an ultrahigh density.
authors.affiliation: Departement Chemie, Katholieke Universiteit Leuven, Celestijnenlaan 200F, B-3001 Heverlee, Belgium. Robert.neely@chem.kuleuven.be
authors.forename: Robert K
authors.initials: RK
authors.lastname: Neely
authors.forename: Jochem
authors.initials: J
authors.lastname: Deen
authors.forename: Johan
authors.initials: J
authors.lastname: Hofkens
mesh_terms: D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004262:DNA Restriction Enzymes; D005456:Fluorescent Dyes; D023281:Genomics / Q000379:methods; D046210:Microfluidic Analytical Techniques; D036103:Nanotechnology; D055620:Optical Phenomena
chemical_list: D005456:Fluorescent Dyes; D004247:DNA; D004262:DNA Restriction Enzymes