250
ANNETTE DEICHMANN AND KLAUS DEICHMANN
• 2 x YT medium (see Subprotocol 3, Materials)
• Chloramphenicol, 30 mg/ml in ethanol, sterile-filtered
• Solution A: 100 mM Tris-Cl (pH 8.0 ), 50 mM N aCl; 15 mM EDT A (pH 8.0)
• Solution B: Add 1.6% Triton X-100 (v/v) to solution A
• Ethidium bromide, 5 mg/ml
• CsCl solution ( 1 g/ml) in TE buffer
• TE buffer: 10 mM Tris; 1 mM EDTA (pH 8.0)
• Isopropanol, saturated with CsCl ( dissolve 15 g CsCl in 15 ml ofTE buffer
and add approximately 100 ml of isopropanol; two phases will appear)
• Solution for dialysis tubing: 2% Na2HC0 3, 1 mM EDTA
• 95% ethanol
• 70% ethanol
• 1 mM EDTA (pH 8.0)
• 3 M Na-acetate (pH 7.0)
Suppliess • Shaker or shaking water bath
• Spectrophotometer
• Centrifuge
• Ultracentrifuge (e.g. Beckman, with VTi50 and VTi65 rotors)
• Quick-seal ultracentrifuge tubes for the selected rotors and sealing apparatus (Beckman)
• UV-lamp
• Dialysis tubing
• Dialysis clips
• Vortex
• Magnetic stirrer
• Eppendorf centrifuge
• Vacuum evaporator
ANNETTE DEICHMANN AND KLAUS DEICHMANN
• 2 x YT medium (see Subprotocol 3, Materials)
• Chloramphenicol, 30 mg/ml in ethanol, sterile-filtered
• Solution A: 100 mM Tris-Cl (pH 8.0 ), 50 mM N aCl; 15 mM EDT A (pH 8.0)
• Solution B: Add 1.6% Triton X-100 (v/v) to solution A
• Ethidium bromide, 5 mg/ml
• CsCl solution ( 1 g/ml) in TE buffer
• TE buffer: 10 mM Tris; 1 mM EDTA (pH 8.0)
• Isopropanol, saturated with CsCl ( dissolve 15 g CsCl in 15 ml ofTE buffer
and add approximately 100 ml of isopropanol; two phases will appear)
• Solution for dialysis tubing: 2% Na2HC0 3, 1 mM EDTA
• 95% ethanol
• 70% ethanol
• 1 mM EDTA (pH 8.0)
• 3 M Na-acetate (pH 7.0)
Suppliess • Shaker or shaking water bath
• Spectrophotometer
• Centrifuge
• Ultracentrifuge (e.g. Beckman, with VTi50 and VTi65 rotors)
• Quick-seal ultracentrifuge tubes for the selected rotors and sealing apparatus (Beckman)
• UV-lamp
• Dialysis tubing
• Dialysis clips
• Vortex
• Magnetic stirrer
• Eppendorf centrifuge
• Vacuum evaporator