African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12072

Full Length Research Paper

A new screening method for selection of desired recombinant plasmids in molecular cloning

  Amirhossein Mohseni, Keivan Majidzadeh-A.* and Mohammad Soleimani            
  Tasnim Biotechnology Research Center (TBRC), AJA University of Medical Science, Tehran, Iran. Academic Center for Culture, Education & Research (ACECR), Iranian Center for Breast Cancer (ICBC), Tehran, Iran.
Email: [email protected]

  •  Accepted: 23 January 2012
  •  Published: 31 August 2012

Abstract

 

One of the problems in cloning process is the low concentration of gene fragment and vector following gel extraction stage which may lead to decreased likelihood of effective ligation. Regarding the facts of this study, after digestion process, the products directly were subjected to ligation. Due to the presence of three antibiotic resistance genes in the reaction, a new strategy based on design of selective media containing different antibiotics was used for selecting the desired colonies. The results of this study indicated that skipping gel extraction process could result in a successful, simple and quick cloning process with high efficiency.

 

Key words: Cloning, gel extraction, vector, screening, antibiotic resistance genes.

Abbreviation

Abbreviation: t-PA, Tissue-type plasminogen activator.

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