Comet Cell Assay 2018

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Summer 2018 Comet Cell Assay.

Objectives

  • Ready to use device (the best would be automatic)
  • Using "kitchen-sink-ingredients" to promote easy to do and safe citizen science

Default Protocol

For the detailed protocol, see Step by step protocol for comet assay


Material needed:

  • Sterile water
  • 0.9% NaCl Saline
  • 1.5% LMP Agarose (made with 0.9% Saline)
  • 0.5M Tris
  • 0.5M EDTA pH 8
  • 5M NaCl
  • 10% Triton
  • H2O2
  • Proteinase K (PK+) solution stock of 25 mg/ml
  • TBE
  • TBE (alkaline)
  • SYBR Safe
  • Electrophoresis machine
  • 12-wells plate


Succinct protocol:

  • Harvest the cells, mix them with agarose to embed them into the gel
  • Once the pads are made, put them in the 12-wells plate, depending on the conditions wanted
  • Wash twice with PK Buffer, then with the Proteinase K
  • Wash twice with alkaline TBE and then twice with normal TBE
  • Run for 18 minutes at 12 V in the Electrophoresis machine
  • Stain with SYBR Safe


Results

Different conditions were tested (...)
Photos (...)