1) Make ~1.1% Low Melting Point agarose gel in 1X TBE. Boil agarose solution, cast gel, and allow to solidify at 4°C.

2) Run sample on gel, do not exceed 60V at room temp so gel does not melt. Stain with weak solution of ethidium bromide

3) Cut out desired band using UV illumination. measure its volume by weight, assume that 1g=1ml.

4) Add 1/10 volume 10x ß-agarase buffer (NEB #392)

5) Heat to 65°C until gel is completely molten, takes about 15 to 30 minutes.

6) Transfer to 40°C, add 1 unit ß-Agarase (NEB#392) per 200µl of gel

7) Incubate 2 hours at 40°C. Digestion of agarose is complete if

solution does not solidify at room temperature.

8) Add sodium acetate to 0.3M, allow to precipitate 10 minutes on ice

9) Spin 15 minutes top speed to pellet undigested carbohydrates

10) Remove supernatant and Phenol /Ø:CHCl3 / CHCl3 Extract

11) Precipitate using standard methods. May want to allow 10-20

minutes for small quantities of DNA to precipitate.

500mM BIS-Tris pH 6.5

100mM EDTA