PLANT DNA MINI-PREP
A slight modification of the Dellaporta technique (Plant Molecular Biology Reporter 1(4): 19

1. Immerse chopped plant material (approximately 2 g) in liquid N2 then grind for 1 min. in a prechilled (with liquid N2) Waring blender.

2. Transfer 1 g of resulting fine powder (still frozen) with a prechilled spatula into a 50 ml Oak Ridge tube containing 16 ml of extraction buffer:

Rx for Mini-prep Extraction Buffer

	To make 100 ml:
	58 ml H2O
100 mM Tris pH 8	10 ml 1 M Tris, pH 8.0
50 mM EDTA pH 8	10 ml 0.5 M EDTA
500 mM NaCl	10 ml 5M NaCl
10 mM b-Mercaptoethanol	70 ml b-Mercaptoethanol
1.25% SDS	12 ml 10% SDS

3. Mix thoroughly by vigorous shaking and place at 65° for 10 minutes.

4. Add 5.0 ml 5 M Potassium acetate. Shake vigorously and incubate at 0° for 20 min.

5. Spin tubes at 15000 rpm for 20 min. in the SS34 rotor. Collect supernatant by pouring through a syringe that has a milipore apparatus attached to it, which contains fine nylon mesh rather than a filter. Collect the supernatant in a Oak Ridge tube containing 10 ml isopropanol. Mix and incubate at -20° C for at least 30 min.

6. Pellet DNA at 15000 rpm for 15 min. Gently pour off supernatant and lightly dry pellets by inverting the tubes on a paper towel for 10 min.

7. Redissolve DNA pellets with 0.7 ml of 50 mM Tris, 10 mM EDTA pH 8, transfer to a microfuge tube, and spin for 10 min. to remove     insoluble debris.

8. Transfer the supernatant to a new microfuge tube. and add 75 ml 3M Sodium acetate and 500 ml isopropanol. Mix well and pellet the clot for 30     seconds in a microfuge. Wash with 80% ethanol, dry and redissolve in 100 ml 10 mM Tris, 1 mM EDTA pH 8.