DNA extract from tissues or cells
- prepare the following lysis buffer (1 liter):
100 mM Tris pH 8.5 (100 ml of 1 M pH8.5 Tris-HCl)
5 mM EDTA (10 ml of 0.5 M EDTA)
0.5% SDS (50 ml of 10% SDS)
200 mM NaCl (40 ml 5M NaCl)
2. add 800 ml water, dissolve it completely.
3. add water to 1 liter.
For DNA extraction, add 1 ml lysis buffer into 1x10^6 cells or equivalent of tissue, mix well, then add proteinase K to the reaction solution at 300 ug/ml.
Incubate it at 55°C for a few hours(cells) or overnight (tissue).
Add 2.5 volume of 95-100% ethanol into the lysis buffer, gently mix. Pick up the white DNA and wash twice in 70% alcohol, air-dry for 10-15 minutes, dissolve into 200-500 ul TE buffer. Store at -4 for future use.
Proteinase K Preparation:
For 100ml of storage buffer:
