Isolation and Purification of DNA from Plants
This chapter describes a DNA extraction method that can be used both on freeze-dried leaves and on fresh leaves, and is based on the method of Saghai-Maroof et al. (1), modified by David Hoisington and Jack Gardiner at the University of Missouri at Columbia (personal communication). The scale of the extraction is dependent on the amount of starting material; 300–400 mg freeze-dried material requires 9 mL extraction buffer and should yield 250 µg-l mg DNA. For high throughput screening, scaling the procedure down tenfold results in a miniprep method that is very suitable for extracting small quantities (25–100 µg) of DNA from young, fresh leaves.
KeywordsFresh Tube Polypropylene Centrifuge Tube Disodium Ethylenediaminetetraacetic Acid Leaf Midrib CTAB Extraction Buffer