Assessment of Various Techniques for Extracting Genomic DNA from Herbarium Specimens of Myosotis

Subject Areas : Plant Biology

zahra ghasemzadeh ¹ , Maryam Khoshsokhan mozaffar ^{2 *}

1 - Ms of Biology, Qo.C. Islamic Azad University, Qom, Iran
2 - Associate Prof. Department of Biology, Qo.C. Islamic Azad University, Qom, Iran

Received: 2025-04-21 Accepted : 2025-05-17 Published : 2025-06-01

Keywords: DNA Extraction, CTAB, Myosotis, Dellaporta, Secondary Metabolites,

Abstract :

Introduction: DNA extraction is a crucial technique in molecular biology. However, extracting DNA from various plants can be challenging due to the presence of secondary metabolites. This study aims to discover an effective method for extracting DNA from Myosotis species, part of the borage family.

Materials and Methods: Three extraction methods were employed on five Myosotis species: unmodified CTAB, modified CTAB, and Dellaporta.

Results: The findings revealed that using unmodified CTAB resulted in a weak band only for M. stricta (collection 2007). In contrast, the modified CTAB method produced strong bands for M. stricta (collections 2007 and 2012) and M. asiatica (collection 2004), while M. sparsiflora (collection 1980) and M. sylvatica (collection 1988) yielded weaker bands. The Dellaporta method showed a band for M. asiatica (collection 2004).

 

Conclusion: The modifications in the CTAB method, which included increasing the concentration of 60% mercaptoethanol (to prevent polyphenol oxidation) and adding ammonium acetate (for improved separation of histones from nucleic acids), resulted in better DNA concentration. Given the potential for DNA degradation in older samples, the modified CTAB method provided superior quality and quantity of extracted DNA.

 

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