Griffitts:Miniprep

Standard Qiagen Miniprep

Materials

• Qiaprep Spin Kit (27106)
• TE buffer

Procedure

1. Grow DH5alpha with appropriate plasmid in liquid LB (4 ml) for 12-18 hours.
2. Spin down 1.3 ml of culture, dump and tap out supernatant, and repeat.
3. Resuspend pellet in 250 ul P1 (from fridge).
4. Add 250 ul P2, immediately invert 3 times, let stand for 2 min.
5. Add 350 ul N3, immediately invert 8 times.
6. Spin 6 min.
7. Carefully move supernatant (~820 ul) to spin column.
8. Spin 30 s.
9. Remove flowthrough and add 650 ul of wash buffer PE to column.
10. Spin 30 s.
11. Remove flowthrough.
12. Spin 60 s.
13. Move column to 1.5 ml tube, add 50-100 ul TE directly to disk, let stand 1-2 min.
14. Spin 60 s to elute DNA.

Adapted from Qiagen handbook