International Journal of
Biotechnology and Molecular Biology Research

  • Abbreviation: Int. J. Biotechnol. Mol. Biol. Res.
  • Language: English
  • ISSN: 2141-2154
  • DOI: 10.5897/IJBMBR
  • Start Year: 2010
  • Published Articles: 100

Genomic DNA extraction protocols from a Moroccan medicinal and aromatic plant Artemisia herba-alba Asso for RAPD-PCR studies

Ourid Ibtissam1,2, Ghanmi Mohamed1, EL Ghadraoui Lahsen2, Kerdouh Benaissa1 and Bakkali Yakhlef Salah Eddine1*
1Centre de Recherche Forestière, Rabat, Marocco. 2Faculté des Sciences et Techniques, Fès, Marocco.  
Email: [email protected]

  •  Accepted: 04 August 2011
  •  Published: 31 August 2011

Abstract

This is the first report on development of protocol for high purity genomic DNA isolation from the Moroccan Artemisia herba-alba schrub leaves and optimization of conditions for RAPD-PCR analysis. Two DNA extraction protocols were specifically developed: QIAgen DNA Kit and protocol developed by Ouenzar et al. (1998). DNA yield and purity were monitored by gel electrophoresis and by determining absorbance at UV (A260/A280). Both ratios were between 1.7 and 2.0, indicating that the presence of contaminating metabolites was minimal. The Ouenzar and collaborators protocol gave higher yield but was more time consuming compared to QIAgen Kit. However, both techniques gave DNA of good quality that is amenable to RAPD-PCR reactions. Additionally, restriction digestion and PCR analyses of the obtained DNA showed its compatibility with downstream applications. Randomly Amplified Polymorphic DNA profiling from the isolated DNA was optimized to produce scorable and clear amplicons. The presented protocols allow easy and high quality DNA isolation for genetic diversity studies within A. herba-alba.

Key words:  Artemisia herba- alba, genomic DNA extraction, PCR-RAPD.