DNA extraction

A B. distachyon leaf was frozen using liquid nitrogen and then ground up.  DNA extraction buffer (DEB) was added.  DEB contains 1% sodium dodecyl sulfate (SDS), a detergent which binds and solubilizes lipids found in the cell membrane.  DEB also contains 25 mM ethylene diamine tetra-acetic acid (EDTA), which binds metal ions that enzymes, such as DNases, require to function.  Potassium acetate (KOAc) was used to precipitate proteins and carbohydrates while nucleic acids remained soluble.  100% isopropanol was used to precipitate the nucleic acids from the salty solution.  The pellet was rinsed with 70% ethanol to remove sodium.  The pellet was resuspended and stored in 50 mL T₁₀E₁ buffer, which contains 10 mM Tris and 1 mM EDTA.