- Remove media with multiple chanel pipette or by aspiration.
- Add 100 µl PBS, remove.
- Add 50 µl lysis buffer.
- Wrap the plate with wet paper towel then seal it in a plastic bag.
- Incubate at 50°C overnight.
- Add 100 µl cold 100 percent etOH.
- Leave at room temp (not shaking).
- If a plate spinner is available, spin plates for 5’ at 1000 rpm at 10°C. Pour off EtOH by blotting on paper towel pad.
- Wash three times with 150 µl 70 percent EtOH and pour off.
- Pat on paper towel pad until very little liquid comes out.
Lysis Buffer
|
Final Concentration |
Stock Solution |
10 ml Lysis Buffer |
| 0.5% Sarcosyl (Sigma L9150) | 10% | 500 µl |
| 200 mM NaCl | 5 M | 400 µl |
| 10 mM EDTA (pH 8.0) | 0.5 M | 200 µl |
| 10 mM Tris.HCl (pH 8.0) | 1 M | 100 µl |
| 1 mg/ml proteinase K | 20 mg | 500 µl |