USER-derived cloning methods and their primer design

Bo Salomonsen, Uffe Hasbro Mortensen, Barbara Ann Halkier

10 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationDNA cloning and assembly methods
Number of pages13
PublisherHumana Press
Publication date2014
Pages59-72
Chapter5
ISBN (Print)978-1-62703-763-1
ISBN (Electronic)978-1-62703-764-8
DOIs
Publication statusPublished - 2014
SeriesMethods in Molecular Biology
Volume1116
ISSN1064-3745

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