High Molecular Weight DNA Extraction of Pomegranate v2

The purpose of this protocol is to share a modified CTAB DNA extraction method that can be generally applied to all plants, but is especially effective in extracting High Molecular Weight (HMW) DNA from plants with high concentrations of secondary plant metabolites. Pomegranates and other plants rich in polyphenolic compounds and polysaccharides, which may have difficult-to-penetrate cell walls, make DNA extraction difficult when the extraction products are meant for long-read sequencing. The most unfortunate scenario resulting from sequencing a plant with high levels of polyphenolics is a partial sequencing run that can be a waste of thousands of dollars. This protocol requires approximately 10 grams of tissue to yield adequate results for HMW extraction. This protocol typically yields approximately 20-40 µg of DNA at a concentration of at least 100ng to an upwards of 1000 ng/µL with a DIN of at least 6.7.