Extracting total genomic DNA of Chrysanthemum sensu lato by CTAB and SDS without both liquid nitrogen and phenol

Abstract

Total genomic DNA was isolated from young leaves of Artemisia arborences L., Achiliea fragrantisma (Forssk.) Sch. Bip, Ach. santolina L., Glebionis coronaria L., G. coronaria var. discolor D' Urv, Cotula barbata DC., Co. cinerea Del, and Matricaria recutita L. of Chrysanhemum senu lato (Asteraceae) as well as an eucalyptus tree as the outgroup by using the CTAB and SDS methods. Three fixative solutions such as alcohol, alcohol-chloroform and alcohol-EDTA were used before grinding the plant leaves to extract the total genomic DNA. The quality and quantity of DNA were comparable among the three fixative solutions and liquid nitrogen grinding after using the CTAB and SDS methods. Isolated DNA was suitable for the ISSR, the RAPD and the 5S rDNA analyses. The CTAB method showed DNA quality much better than SDS. This method did not require liquid nitrogen for fixation, grinding, or storage at -196°C as well as phenols for washing after the CTAB and SDS DNA extraction, making the present method advantageous and more suitable in comparison with common protocols for the plant materials in Chrysanhemum senu lato.