Background: The manuscript is originally the internship report of Muhammad Junaid, submitted for the partial fulfilment of the degree of BSc (h) to University of Swabi. The internship was completed at NIFA (Nuclear Institute of Food and Agriculture), Peshawar, KP-Pakistan under the supervision of Syed Tariq Shah, Senior Scientist, NIFA Peshawar in 2014. The results obtained are from the original project of Syed Tariq Shah, Senior Scientist, NIFA Peshawar. Methodology: Ten advance lines of wheat were surface sterilized by 70% ethanol for two minutes and then rinse with double distilled water three times. Again washed with mercuric chloride and washed with double distilled water for three times. The sterilized seeds were cultured in Petri dish and kept it in growth chamber at 25oC. When the plants achieved optimum size after germination, the flag leaf were collected on the basis of weight and stored at -80 for molecular analysis. Genomic DNA was extracted from these leaves using DNeasy plant mini kit. Agarose gel was run after the DNA extraction and then the Spectrophotometric Determination was done. Results: The results literally focused on to study an efficient method for extraction of genomic DNA and to determine the quality and quantity of genomic DNA. After the extraction of genomic DNA from the plant tissues the quantification, quality and concentrations of isolated genomic DNA were verified on 1% of Agarose gel. RNA and other polysaccharides proteins contamination in the Data is not recoded providing a very rapid and sensitive means of estimating the nucleic acid concentration. DNA concentration was also measured and quantified by ultraviolet spectrophotometer using ultraviolet spectrophotometer. Results showed the differences in DNA yield and DNA quality among the advanced lines of ten wheat samples. Purified genomic DNA has A260/A280 ratios of 1.5–1.77, and absorbance confirming high purity. L-10 shows the highest yield of DNA that is 462.47 and the lowest DNA yield is of L-7 that is 355.54, the OD260/280 of L-3 is high then the remaining nine lines. Conclusion: DNeasy plant mini kit is an efficient for genomic DNA extraction and purification.