Large scale purification of linear plasmid DNA for efficient high throughput cloning.
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.
Noirclerc-Savoye
Marjolaine
M
CEA, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France. marjolaine.noirclerc@ibs.fr
Gallet
Benoit
B
Bernaudat
Florent
F
Vernet
Thierry
T
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
D008565:Membrane Proteins; D012117:Resins, Synthetic; C042855:Mono Q