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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.

SUBMITTER: Fujishima K

PROVIDER: S-EPMC4443763 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature

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An overhang-based DNA block shuffling method for creating a customized random library.

Fujishima Kosuke K Venter Chris C Wang Kendrick K Ferreira Raphael R Rothschild Lynn J LJ

Scientific reports 20150526

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 an ...[more]

PMID: 26010273

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