Genomic DNA extraction from mouse tail by proteinase K
Genomic DNA extraction from mouse tail by proteinase K
Material
- DNA lysis buf (100 mM Tris-HCl pH 8.5/5 mM EDTA/0.2% SDS/200 mM NaCl, RT)
- Proteinase K (20 mg/ml stock, -20C) (Clontech 740506, 4C)
Day 1
- Cut 5-10 mm mouse tail and put it into 1.5 ml tube.
- Add 500 ul DNA lysis buf with 40 ug/ml proteinase K (1/500 of stock).
- Incubate at 55C for o/n.
Day 2
- Add 500 ul phenol.
- Mix by tube mixer for 5 min.
- Centrifuge at 14000 rpm at RT for 10 min.
- Move supernatant to new tube.
- Add 500 ul chloroform.
- Mix by tube mixer for 5 min.
- Centrifuge at 14000 rpm at RT for 10 min.
- Move supernatant to new 1.5 ml tube.
- Add 500 ul isopropanol.
- Shake to mix (gDNA can be seen as white precipitate).
- Centrifuge at 14000 rpm at RT for 10 min.
- Remove supernatant by decantation.
- Add 500 ul of 70% EtOH.
- Centrifuge at 14000 rpm at RT for 5 min.
- Remove supernatant by 200 ul pipet.
- Dry up.
- Dissolve in 100 ul H2O.
- Use 1 ul for PCR template.
