Preparation of Tail Samples (for Genotyping)
PBND Solution: Tail Lysis Buffer
| Reagent | Final Concentration | From Stock (for 250 mL) |
|---|---|---|
| Tris pH 8.0 | 10 mM | 2.5 mL of 1M |
| KCl | 50 mM | 12.5 mL of 1M |
| MgCl2 | 0.1 mg/mL | 260 uL of 1M |
| NP40 | 0.45% | 11.25 mL of 10% |
| Tween 20 | 0.45% | 11.25 mL of 10% |
| Gelatin | 0.1 mg/mL | 25 mg |
Add the above to ~200 mL of water, check that pH is 8.3 and then bring up to 250 mL. Store at 4 degrees
For each digestion, add 2 uL of 10 mg/mL proteinase K diluted in 50 uL PBND. Combine as a master mix and then aliquot into wells with tissue.
Protocol
- Combine 100uL of PBND solution with 2ul of Proteinase K and mouse tail in an eppendorf tube or in a well of a 96 well PCR plate (Proteinase K stock = 10mg/mL in ddH2O)
- Incubate at 55 degrees (16 hours - O/N)
- Incubate at 85 degrees for 60 min
- Hold at 4 degrees
If you need higher purity DNA try the protocol at DNA Preparation from Tail Clip