title: A modified version of the digestion-ligation cloning method for more efficient molecular cloning.
abstract: Here we describe a modified version of the digestion-ligation approach for efficient molecular cloning. In comparison with the original method, the modified method has the additional steps of gel purification and a second ligation after the first ligation of the linearized vector and DNA insert. During this process, the efficiency and reproducibility could be significantly improved for both stick-end cloning and blunt-end cloning. As an improvement of the very important molecular cloning technique, this method may find a wide range of applications in bioscience and biotechnology.
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Jiannan
authors.initials: J
authors.lastname: Zhang
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Hongman
authors.initials: H
authors.lastname: Chen
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Daming
authors.initials: D
authors.lastname: Ren
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhang
authors.affiliation: College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China. Electronic address: anyingfeng666@163.com.
authors.forename: Yingfeng
authors.initials: Y
authors.lastname: An
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004247:DNA; D004262:DNA Restriction Enzymes; D011088:DNA Ligases
keywords: Blunt-end DNA ligation; Digestion–ligation approach; Molecular cloning; T4 DNA ligase