title: Cloning technologies.
abstract: One major obstacle in realizing the potential behind human embryonic stem cells (hESC) is the availability of efficient and reliable engineering methods. Such methods require cloning technologies that can be applied to a variety of platforms and can serve multiple functions. In the last two decades cloning technologies have become more efficient, widening the bottleneck in creating engineered hESC lines. Using TOPO(®) TA cloning kits, genes can be efficiently amplified and inserted into target vectors with minimal manipulation and purification. For more complex cloning procedures we introduce the Multisite Gateway(®) system. This is a cloning platform based on integrase technology that allows for the generation of complex multicistronic gene configurations that can transverse a variety of platforms with ease. These technologies allow the end user to quickly and efficiently select clones, as well as combine multiple genetic elements of interest between platform technologies in a high-throughput manner, providing scientists with a toolbox to create tools to dissect stem cell biology.
authors.affiliation: Primary and Stem Cell Systems, Life Technologies, Carlsbad, CA, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Fontes
mesh_terms: D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000502:physiology*; D004926:Escherichia coli; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers