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Seamless Ligation Cloning Extract (SLiCE) cloning method.

Methods in molecular biology (Clifton, N.J.) (2014-01-08)
Yongwei Zhang, Uwe Werling, Winfried Edelmann
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-(+)-Arabinose, ≥99% (GC)
Sigma-Aldrich
CelLytic B Cell Lysis Reagent, For bacterial cell lysis, standard strength