IGEM:Harvard/2006/vlau/Notebook/2006-7-13: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
|||
Line 40: | Line 40: | ||
- 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. | ||
Latest revision as of 09:16, 2 August 2006
Protein & DNA-staining Experiment (Repeat 1)
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 | 8 | 0.65 | 0.5 | 6.5 | 3.0 | 10.0 |
2 | 9 | 1.30 | 1.0 | 6.0 | 3.0 | 10.0 |
3 | 10 | 1.95 | 1.5 | 5.5 | 3.0 | 10.0 |
4 | 11 | 2.69 | 2.0 | 5.0 | 3.0 | 10.0 |
Tube | Lane | 2 μM Aptamer (μL) | water (μL) | 10x loading dye (μL) | total volume (μL) |
1 | 2 | 1.0 | 6.0 | 3.0 | 10.0 |
2 | 3 | 2.0 | 5.0 | 3.0 | 10.0 |
3 | 4 | 3.0 | 4.0 | 3.0 | 10.0 |
4 | 5 | 4.0 | 3.0 | 3.0 | 10.0 |
2. Protocol
- gel run @ 120V for 1.25 hrs. - 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.