Evaluation of seamless ligation cloning extract preparation methods from an Escherichia coli laboratory strain

Yuki Okegawa, Ken Motohashi

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

Seamless ligation cloning extract (SLiCE) is a simple and efficient method for DNA cloning without the use of restriction enzymes. Instead, SLiCE uses homologous recombination activities from Escherichia coli cell lysates. To date, SLiCE preparation has been performed using an expensive commercially available lytic reagent. To expand the utility of the SLiCE method, we evaluated different methods for SLiCE preparation that avoid using this reagent. Consequently, cell extracts prepared with buffers containing Triton X-100, which is a common and low-cost nonionic detergent, exhibited sufficient cloning activity for seamless gene incorporation into a vector.

Original languageEnglish
Pages (from-to)51-53
Number of pages3
JournalAnalytical Biochemistry
Volume486
DOIs
Publication statusPublished - Jul 27 2015
Externally publishedYes

Keywords

  • Detergent extraction
  • Homologous recombination
  • Seamless DNA cloning
  • SLiCE

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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