Difference between revisions of "Comet Cell Assay 2018"
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* Using "[[kitchen-sink-ingredients]]" to promote easy to do and safe citizen science | * Using "[[kitchen-sink-ingredients]]" to promote easy to do and safe citizen science | ||
| − | ==Protocol== | + | ==Default Protocol== |
''For the detailed protocol, see [[Step by step protocol for comet assay]]'' | ''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 (...) <br> | ||
| + | Photos (...) | ||
| + | |||
| + | [[Category:Biology protocols]] | ||
| + | [[Category:Documentation]] | ||
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 (...)