RNA Isolation

  1. Dielectrophoresis treatment solutions.
  2. Add 0.3 mL lysing buffer (0.1 M HEPES, 0.2 M EDTA, 10% SDS, 0.02 M
    EGTA (pH 7.5) [DEP-treated) to 3 mL bacterial solution.
  3. Extract with 1 vol 1000:140:0.4 phenol:m-cresol:hydroxyquinoline.
  4. Add directly to 2 volumes ethanol (=6.6 mL).
  5. Precipitate for 16 hrs at –20°C.

RNA Preparation and Electrophoresis
Buffers needed:
  1. 0.2 M MOPS (pH 7.0), 50 mM sodium acetate, 1 mM EDTA.
  2. Formaldehyde, 37%.
  3. Formamide—deionized until pH is 7.0 and recrystallized at 0°C, stored at –20°C.
  4. 50% glycerol, 1 mM EDTA, 0.4% bromphenol blue, 0.4% xylene cyanol.

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