Abstract
Aims: Artemisia judaica in Saudi Arabia are exposed to serious threats due to heavy human impacts such as uncontrolled tourism, overgrazing and uncontrolled collection, mining, quarrying and natural drought. These activities warrant careful conservation strategies that would allow inclusion of these plants into development plans of the Saudi Arabia economy in order to ensure their sustainable use. The evaluation of the genetic diversity by using ISSR and RAPD PCR requires a high quality of intact DNA. Therefore the aim of this study was to establish a rapid DNA isolation protocol for Artemisia species without using Liquid Nitrogen.
Results: In case of Artemisia species, the DNA protocol was optimized by using solution based method without using Liquid Nitrogen. The quality of DNA was checked on 0.7% agarose gel containing ethidium bromide at a final concentration of 0.5 mg/ml. This provides a very rapid and sensitive means of estimating the nucleic acid concentration. Similarly, the quantity and purity of DNA extracted from Artemisia species analyzed by using NanoDrop1000. The lambda 260/lambda 280 determined for Artemisia DNA samples were in a range of 1.83-2.0 and concentration (ug/ml) (502.7 to 1288.5 ng/mu l) indicating a good quality DNA from our method. The protocol was simple, reliable and operative in normal laboratory condition in two hours.
Conclusion: The developed protocol proved to be simple, reliable and operative in normal laboratory condition yielding intact DNA from Artemisia species having good quantity (170 to 1200 ng/mu l) and quality (A(260/280) ratio - 1.64 to 1.95) completed in two 2 to 3 hours.