IGEM:Harvard/2006/vlau/Notebook/2006-7-12

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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)
110.650.58.52.011.0
221.301.08.02.011.0
332.602.07.02.011.0
443.903.06.02.011.0
555.204.05.02.011.0
Tube Lane 2 μM Aptamer (μL) water (μL) 10x loading dye (μL) total volume (μL)
171.08.03.012.0
282.07.03.012.0
393.06.03.012.0
4104.05.03.012.0
5115.04.03.012.0

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?
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