Kun:LabNotes/Haplotyping/Protocols
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Lysing mammalian cells[edit]
- Reagents:
Lysis buffer 0.45% Tween, 0.45% NP40, 2.5mM MgCl2, 50mM KCl, 10mM Tris-Cl pH8.3, 100ug/ml Proteinase K
Tween20 225ul NP40 225ul 1M MgCl2 125ul 1M KCl 2.5ml 1M Tris-Cl pH8.3 0.5ml 20ug/ul Prot K 250ul* dH2O 46.2ml * Add Proteinase K just before use, 5ul stock per ml of lysis buffer used.
- Protocol:
- Wash cells with cold PBS.
- Resuspend at 10000 cells/ul in lysis buffer.
- Incubate at 56 C for 1 hour.
- Boil for 10 minutes.
- Store lysate at -20 C
- Another lysis buffer recipe
20 mM EDTA 10 mM Tris 8.0 200 mM NaCl 0.2% Triton X-100 100 mg/ml PK Incubate at 37C for 1.5 hour.