SPERA Assay

Procedure

  1. Grow saturated culture of Klebsiella.
  2. Spin down and resuspend in 1/4 voume HL-5 (no drugs).
  3. Apply to Sm/16 plate by adding 1ml bacterial suspension to 2-3 days old plate. Tilt and rotate plate to cover entire surface. Tilt excess into edge and aspirate off.
  4. Incubate overnight .
  5. Harvest dicty containing plasmid: Use 3ml from a confluent 10mL (100mm) plate. Spin (4min/1500rpm) and resuspend in 100ul undrugged HL-5.
  6. Apply Dicty to lawn by carefully dripping 8ul/drop onto plate. Plate 2-6 drops per dicty type.
  7. Incubate at 21C o/n..
  8. Measure diameter of plaques over time-points.