Seamless Ligation Cloning Extract (SLiCE) cloning method

Research output: Chapter in Book/Report/Conference proceedingChapter

23 Scopus citations

Abstract

SLiCE (Seamless Ligation Cloning Extract) is a novel cloning method that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (15-52 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from bacterial artificial chromosomes or other sources. SLiCE is highly cost-effective and demonstrates the versatility as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. We established a DH10B-derived E. coli strain expressing an optimized λ prophage Red recombination system, termed PPY, which facilitates SLiCE with very high efficiencies.

Original languageEnglish (US)
Title of host publicationDNA Cloning and Assembly Methods
PublisherHumana Press Inc.
Pages235-244
Number of pages10
ISBN (Print)9781627037631
DOIs
StatePublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1116
ISSN (Print)1064-3745

Keywords

  • Bacterial cell extract
  • DNA cloning
  • In vitro recombination
  • Recombinant DNA
  • SLiCE cloning
  • Seamless cloning

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Zhang, Y., Werling, U., & Edelmann, W. (2014). Seamless Ligation Cloning Extract (SLiCE) cloning method. In DNA Cloning and Assembly Methods (pp. 235-244). (Methods in Molecular Biology; Vol. 1116). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-764-8_16