A simple and efficient DNA extraction protocol for old herbarium leaves of Bellevalia (Asparagaceae, Scilloideae)

Jalali Roudsary, Lida; Jafari, Azarnoosh; Vaezi, Jamil; Karimi, Ehsan

doi:10.52547/nbr.9.2.124

A simple and efficient DNA extraction protocol for old herbarium leaves of Bellevalia (Asparagaceae, Scilloideae)

Authors

Lida Jalali Roudsary ¹

Azarnoosh Jafari ¹

Jamil Vaezi ²

Ehsan Karimi ¹

¹ Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran

² Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran; Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

https://doi.org/10.52547/nbr.9.2.124

Abstract

High-quality DNA extraction plays an important role to make sharp bands in the gel electrophoresis and also produces clean chromatograms. Usually, DNA extract is delivered using the modified CTAB method but this method cannot obtain high-quality DNA for molecular analysis from old dried leaves of Bellevalia due to having different chemical compounds which inhibit to obtain a clear DNA extraction. To solve the problem, in different phases of a modified CTAB protocol, activated carbon, phenol, potassium acetate, RNase and deionized water (instead of TE) were applied to eliminate contaminants and improve obtained high-quality DNA. The DNA absorption data from the modified method were compared with CTAB method in 260 and 280 nm wavelengths. The results showed a 15-80-fold change in the DNA concentration. This study suggests the modified method of extraction is more efficient in the quality of DNA obtained from the little amount of herbarium old leaves of Bellevalia than that of the CTAB method.

Keywords

activated charcoal

Bellevalia

DNA purification

Phenol

nanodrop

20.1001.1.24236330.2022.9.2.4.6

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