Instruction for the Invisorb Spin Cell Mini Kit

Protocol 1: DNA isolation from cell culture (24-Well-Plates;max. 2x10 6 cells)Important: Incubate the Elution Buffer D at 70°C1. Remove medium and wash cells twice with 1 x PBS (not included in the kit).2. Add 500 Al Lysis Buffer D per well, pipet up and down several times and incubate at roomtemperature for 3 min.Note:Should Lysis Buffer D contain any precipitates incubate it in a 50°C waterbath for a short time!3. Transfer the whole cell lysis suspension into a 1.5 ml or 2.0 ml microcentrifuge tube.Note:Vortex the Carrier Suspension B thoroughly prior to use.4. Addition of 10 Al Carrier Suspension B and 200 Al Binding Buffer HL, mix briefly andtransfer the suspension directly onto the RTA Spin Filter. Incubation for 2 min.5. Centrifugation for 3 min at 13.400 x g (12.000 rpm). Discard the flow-through and reuse theRTA Receiver Tube.6. Add 550 Al Wash Buffer and centrifuge for 1 min at 13.400 x g (12.000 rpm). Discard theflow-through.7. Repeat step 6 once again. Afterwards centrifuge for 3 min at 13.400 x g (12.000 rpm) toremove the residual ethanol.8. Transfer the RTA Spin Filter into a new 1.5 ml Receiver Tube. Add 200 Al of ElutionBuffer D or Tris buffered ddH 2 0 (pH 8.5 - 9.0) prewarmed to 70°C directly onto themembrane of the RTA Spin Filter.Incubation for 2 min. Centrifugation at 9.300 x g (10.000 rpm) for 2 min.Discard the RTA Spin Filter.12Invisorb ® Spin Cell Mini Kit 032010