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Large scale purification of linear plasmid DNA for efficient high throughput cloning
Author(s) -
NoirclercSavoye Marjolaine,
Gallet Benoit,
Bernaudat Florent,
Vernet Thierry
Publication year - 2010
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201000132
Subject(s) - cloning (programming) , plasmid , restriction enzyme , dna , cloning vector , ligation , molecular cloning , throughput , restriction digest , computational biology , gene , vector (molecular biology) , biology , microbiology and biotechnology , chemistry , recombinant dna , genetics , computer science , gene expression , telecommunications , wireless , programming language
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(tm) technique confirmed the high quality of the linearized vector as 100% of the genes were successfully cloned.