title: USER-derived cloning methods and their primer design.
abstract: Uracil excision-based cloning through USER™ (Uracil-Specific Excision Reagent) is an efficient ligase-free cloning technique that comprises USER cloning, USER fusion, and USER cassette-free (UCF) USER fusion. These USER-derived cloning techniques enable seamless assembly of multiple DNA fragments in one construct. Though governed by a few simple rules primer design for USER-based fusion of PCR fragments can prove time-consuming for inexperienced users. The Primer Help for USER (PHUSER) software is an easy-to-use primer design tool for USER-based methods. In this chapter, we present a PHUSER software protocol for designing primers for USER-derived cloning techniques.
authors.affiliation: DynaMo Center of Excellence, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Bo
authors.initials: B
authors.lastname: Salomonsen
authors.forename: Uffe H
authors.initials: UH
authors.lastname: Mortensen
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Halkier
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012984:Software
chemical_list: D017931:DNA Primers