IGEM:Harvard/2006/vlau/Notebook/2006-7-12: Difference between revisions
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- protein section stained w/ GelCode Blue Stain (Coomassie Blue) for 1 hr. | - protein section stained w/ GelCode Blue Stain (Coomassie Blue) for 1 hr. | ||
- DNA section stained w/ 10{{ul}} EtBr in 100 mL Tris-glycine buffer for 1 hr. | - DNA section stained w/ 10{{ul}} EtBr in 100 mL Tris-glycine buffer for 1 hr. | ||
3. Results | |||
- both sections showed no bands | |||
- possible reasons: buffer? the gel itself? | |||
Revision as of 13:36, 12 July 2006
Protein & DNA-staining Experiment
1. Rxn Mixtures
| Tube | Lane | Thrombin mass (μg) | 2 μM thrombin (μL) (2 μM = 1.299 μg/μL) |
water (μL) | 10x loading dye (μL) | total volume (μL) |
| 1 | 1 | 0.65 | 0.5 | 8.5 | 2.0 | 11.0 |
| 2 | 2 | 1.3 | 1.0 | 8.0 | 2.0 | 11.0 |
| 3 | 3 | 2.6 | 2.0 | 7.0 | 2.0 | 11.0 |
| 4 | 4 | 3.9 | 3.0 | 6.0 | 2.0 | 11.0 |
| 5 | 5 | 5.2 | 4.0 | 5.0 | 2.0 | 11.0 |
| Tube | Lane | 2 μM Aptamer (μL) | water (μL) | 10x loading dye (μL) | total volume (μL) |
| 1 | 7 | 1.0 | 8.0 | 3.0 | 12.0 |
| 2 | 8 | 2.0 | 7.0 | 3.0 | 12.0 |
| 3 | 9 | 3.0 | 6.0 | 3.0 | 12.0 |
| 4 | 10 | 4.0 | 5.0 | 3.0 | 12.0 |
| 5 | 11 | 5.0 | 4.0 | 3.0 | 12.0 |
2. Protocol
- gel run @ 25V, 4C for 2.5 hrs. (dye appears to have diffused sideways rather than traveling downwards) - gel cut into DNA & protein sections after removal from cartridge (slight tearing towards bottom) - protein section stained w/ GelCode Blue Stain (Coomassie Blue) for 1 hr. - DNA section stained w/ 10μL EtBr in 100 mL Tris-glycine buffer for 1 hr.
3. Results
- both sections showed no bands - possible reasons: buffer? the gel itself?
