Seamless Ligation Cloning Extract (SLiCE) cloning method

Research output: Chapter in Book/Report/Conference proceedingChapter

17 Citations (Scopus)

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 publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages235-244
Number of pages10
Volume1116
ISBN (Print)9781627037631
DOIs
StatePublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1116
ISSN (Print)10643745

Fingerprint

Ligation
Organism Cloning
Genetic Recombination
Prophages
Bacterial Artificial Chromosomes
Recombinant DNA
DNA
Cell Extracts
Escherichia coli
Costs and Cost Analysis

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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

Seamless Ligation Cloning Extract (SLiCE) cloning method. / Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried.

Methods in Molecular Biology. Vol. 1116 Humana Press Inc., 2014. p. 235-244 (Methods in Molecular Biology; Vol. 1116).

Research output: Chapter in Book/Report/Conference proceedingChapter

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