1. Rinse tubes with distilled water and centrifuge the DNA stock to remove any debris which might later clog your injection needle.

2. Add 10 ug of plasmid to 300 ul of TE/0.1 M NaCl.

3. Add 15 ul of 100 mM spermine and ice for 15'.

4. Microfuge in the cold for 15' and discard the supernatant.

5. Dissolve the precipitate in 100 ul of 0.5 M NaCl.  The DNA dissolves slowly so intermittently agitate the tube over a period of 10 minutes.

6. Add 100 ul of water and 400 ul of ethanol.  Ice for 15'.

7. Microfuge in the cold for 15' and discard the supernatant.

8. Dissolve the DNA in 50 ul of water and then add 50 ul of 0.5 M NaCl.

9. Add 200 ul of ethanol, mix, ice 15', microfuge 15', and discard supernatant.

10. Rinse DNA precipitate with 75% ethanol, microfuge 5' and discard supernatant.

11. Air dry precipitate and then dissolve the DNA in 10 ul of injection buffer (0.1 mM Sodium Phosphate pH 6.8, 5 mM KCl).

12. Quantify DNA and adjust to 500 ng/ul with additional injection buffer.