Yu:quick miniprep

Procedure

1. Spin down 1-3 mL culture
   1. 4000 RPM for 5min at 4°C
2. Transfer pellet to eppendorf tube if not already in one
3. Resuspend pellet in 200 μL P1
4. Add 200 μL of P2
   1. Mix by inversion - solution will become very viscous
5. Add 200 μL of P3
   1. Mix by inversion - white precipitate will form
6. Spin mixture for 10 min at top speed
7. Remove pellet with a sterile toothpick
8. Add 600 μL Isopropanol
9. Spin mixture for 3 min at top speed
   1. Decant supernatant
10. Add 500 µl 70% EtOH and vortex briefly
11. Spin mixture for 3 min at top speed
    1. Decant supernatant
12. Air dry pellet
13. Resuspend pellet if 50-100 μL TE₈

Solutions

P1

Final Concentration	For 50 mL	For 100 mL
50 mM Tris pH8.0	2.5 mL of 1M Tris pH8	5 mL of 1M Tris pH8
20 mM EDTA	2.0 mL 0f 0.5 M EDTA	4.0 mL 0f 0.5 M EDTA
milliQ H2O	to 50 mL	to 100 mL

P2

Final Concentration	For 50 mL	For 100 mL
0.2N NaOH	1 mL of 10 N NaOH	2 mL of 10 N NaOH
1% SDS	2.5 mL 0f 20% SDS	5 mL 0f 20% SDS
milliQ H2O	to 50 mL	to 100 mL

P3

3M NaOAc pH4.8