Isolation of high-quality DNA from cotton and its fungal pathogens

A single-seed DNA extraction method was developed to extract high quality complex genomic DNA from different cotton tissues (leaves and seeds) as well as from mycelium. of its fungal pathogens. Seeds were germinated in 24-well cell culture cluster and 5-days-old seedling tissue was used to extract DNA. The average yield of extracted DNA varied from 50 to 100 ng of DNA per 100 mg of processed tissue, allowing over 150 PCR experiments. To investigate the effectiveness of our protocol, DNA extracts were subjected to different treatments, including restriction enzyme digestion, RAPD and ITS-PCR amplification. in all cases, DNA suitable for PCR and for restriction was prepared in less than 60 min. The procedure described here works well for extracting high-quality DNA from cotton tissue and should be widely applicable for an analysis of large populations from all cotton genotypes and its fungal pathogens. The method does not require expensive and environmentally hazardous reagents and equipment. It can be performed even in low-technology laboratories.