African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Short Communication

Blunt-end vectors generated by polymerase chain reaction (PCR) for direct cloning of blunt-end DNA fragments

Dongming Lan1, Huiling Yan2, Bo Yang1 and Yonghua Wang2*
  1School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China. 2College of Light Industry and Food Sciences, Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou 510641, China.
Email: [email protected]

  •  Accepted: 23 February 2011
  •  Published: 19 September 2011

Abstract

 

Blunt-end cloning is a convenient way to clone polymerase chain reaction (PCR) products generated by proof-reading DNA polymerase. However, it is a time consuming procedure to prepare the linearized blunt-end vector, which usually involves plasmid extraction and restriction enzyme digestion. Moreover, 5’ dephosporylation of the vector is usually required to avoid vector self-ligation. Here, we reported a method for generating linearized blunt-end vector pBSK-blunt by PCR. Vector generated in this way has no 5’-phosphate groups, hence completely avoiding vector self-ligation and yielding almost 100% positive clones.

 

Key words: Blunt-end cloning, phosphorylated DNA fragment, dephosphorylated blunt-end vector.

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This article is published under the terms of the Creative Commons Attribution License 4.0

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