Abstract: Two methods were evaluated for the extraction of DNA from barley (Hordeum vulgare L.). A CTAB-based method and a nuclei isolation and extraction method. Results showed that barley DNA extracted by nuclei genomic DNA method was of a high molecular weight (HMW) with a very slight shearing. Whereas, DNA extracted by CTAB method gave a high molecular weight (HMW) with the heaviest DNA shearing. All nuclei DNA extracted samples did not show RNA contamination. However, all CTAB extracted samples, showed RNA contamination. Results obtained from the gel of Pulse Field Gel Electrophoresis (PFGE) and Agarose gel electrophoresis were almost similar except for High-LoTM DNA marker (60-10000 bp) that did not appeared in PFGE due to the long period of the run. Furthermore, the two DNA extraction procedures seemed to be simple and efficient in extracting genomic DNA. But, the nuclei method is more effective in extracting intact and HMW of genomic DNA.