title: An overhang-based DNA block shuffling method for creating a customized random library.
abstract: We present an overhang-based DNA block shuffling method to create a customized random DNA library with flexible sequence design and length. Our method enables the efficient and seamless assembly of short DNA blocks with dinucleotide overhangs through a simple ligation process. Next generation sequencing analysis of the assembled DNA library revealed that ligation was accurate, directional and unbiased. This straightforward DNA assembly method should fulfill the versatile needs of both in vivo and in vitro functional screening of random peptides and RNA created with a desired amino acid and nucleotide composition, as well as making highly repetitive gene constructs that are difficult to synthesize de novo.
authors.affiliation: University Affiliated Research Center, NASA Ames Research Center, Moffett Field, CA, USA.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Fujishima
authors.affiliation: NASA Education Associates Program (EAP), NASA Ames Research Center, Moffett Field, CA, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Venter
authors.affiliation: 1] NASA Education Associates Program (EAP), NASA Ames Research Center, Moffett Field, CA, USA [2] Department of Bioengineering, Stanford University, Stanford, CA, USA.
authors.forename: Kendrick
authors.initials: K
authors.lastname: Wang
authors.affiliation: Université Paris VII - Diderot, France.
authors.forename: Raphael
authors.initials: R
authors.lastname: Ferreira
authors.affiliation: NASA Ames Research Center, Moffett Field, CA, USA.
authors.forename: Lynn J
authors.initials: LJ
authors.lastname: Rothschild
mesh_terms: D018389:Codon, Nonsense; D004247:DNA / Q000737:chemistry*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing; D020125:Mutation, Missense; D017422:Sequence Analysis, DNA
chemical_list: D018389:Codon, Nonsense; D004247:DNA