Shauns Miniprep Protocol

Instructions

1. Vortex culture briefly and pour ~1.5mL into eppendorf tube.
2. Spin @ max for 1 minutes.
3. Aspirate media ­ be careful not to suck up pellet.
4. Resuspend pellet in 250uL of Solution 1
5. Add 250uL of Solution 2.  Cap and invert to mix ~ 8 times.
6. Add 250uL of Solution 3.  Cap and invert to mix ~ 8 times.
7. Spin @ max for 15 minutes.
8. Transfer supernatant to new tube.  (~750uL)
9. Add 750uL 100% Isopropanol.  Vortex and spin for 1 min @ max
10. Aspirate supernatant.
11. Wash w/ 750uL 70% EtOH.  Vortex briefly.
12. Spin @ max for 1 min.
13. Aspirate supernatant.
14. Uncap and allow tubes to dry for 5 minutes.
15. Add 50uL TE w/RNAse added.

Solutions

Solution 1

• 50mM Glucose
• 25mMN Tris-Cl pH 8.0
• 10mM EDTA pH 8.0

Solution 2

• 0.2g NaOH ~ 1 nugget
• 2.5mL 10% SDS
• 22.5mL ddH20

Solution 3

• 60.0mL 5M Potassium Acetate
• 11.5mL Glacial Acetic Acid
• 28.5mL ddH20