title: Large scale purification of linear plasmid DNA for efficient high throughput cloning.
abstract: In this report we describe a rapid, simple, and efficient method for large-scale purification of linear plasmid DNA to answer demand from high-throughput gene cloning. The process is based on the separation of the linear vector from small DNA fragments by anion exchange chromatography. Gene cloning experiments by restriction/ligation or the In-Fusion technique confirmed the high quality of the linearized vector as 100% of the genes were successfully cloned.
authors.affiliation: CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France. marjolaine.noirclerc@ibs.fr
authors.forename: Marjolaine
authors.initials: M
authors.lastname: Noirclerc-Savoye
authors.forename: Benoit
authors.initials: B
authors.lastname: Gallet
authors.forename: Florent
authors.initials: F
authors.lastname: Bernaudat
authors.forename: Thierry
authors.initials: T
authors.lastname: Vernet
mesh_terms: D002852:Chromatography, Ion Exchange / Q000379:methods*; D003001:Cloning, Molecular; D053938:DNA Fragmentation; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000302:isolation & purification*; D012117:Resins, Synthetic
chemical_list: D008565:Membrane Proteins; D012117:Resins, Synthetic; C042855:Mono Q