Comet Cell Assay 2018
Jump to navigation
Jump to search
The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
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 (...)