Comet Cell Assay 2018

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 (...)