PHENOL-CHLOROFORM EXTRACTION

1. to tubes containing DNA, add equal volume of phenol-chloroform

2. mix (invert several times and microfuge for 2 minutes)

3. remove top phase (sample) to tube containing equal volume phenol-chloroform

4. mix (invert several times and microfuge for 2 minutes)

5. remove top phase (sample) to tube containing equal volume chloroform

6. mix (invert several times and microfuge for 2 minutes)

7. remove top phase to fresh tube

8. add 1/10 volume LiCl (or Na⁺Acetate)

9. add 3x volume EtOH; incubate 10 minutes on ice; spin 10 minutes in cold; remove EtOH

10. add 1 ml 80% EtOH; spin 10 minutes in cold; remove EtOH; dry pellet

11. re-suspend in TE (appropriate volume according to amount of DNA)