Protocol
- Cut tail pieces (3mm) and mark ears.
- Add 720 ml STE and 30 ml Proteinase K (10 mg/ml stock).
- Incubate at 55°C on heating block and vortex every hour for 3 hours at top speed for 10 seconds.
- Inactivate proteinase K at 70°C for 5 minutes.
- Quench on ice for 5 minutes.
- Centrifuge tail DNA for 10 minutes at full speed.
- Decant into new tube containing 720 ml Isopropanol.
- Precipitate DNA by inverting the tube or vortexing.
- Spin down DNA for 5 minutes at full speed. Remove supernatant.
- Wash pellet with 70% ethanol.
- Spin down genomic DNA 5 minutes at full speed. Remove supernatant.
- Allow DNA to dry for 1-2 minutes.
- Resuspend DNA in 100-200 ml depending on size of pellet.
- Place tube at 55°C for 1 hour to facilitate dissolution of DNA.
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| STE | Buffer | 100 mm Tris; pH 8.5 / 5 mM EDTA / 0.2% SDS / 200 mM NaCl / in H2O | ||
| TE | Buffer | 100 mM Tris; pH 5.0 / 1 mM EDTA / 10 mM NaCl |
Cite This Article
Zangala, T. Isolation of Genomic DNA from Mouse Tails. J. Vis. Exp. (6), e246, doi:10.3791/246 (2007).