Difference between revisions of "Comet Cell Assay 2018"
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Latest revision as of 14:33, 12 November 2026
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 (...)