Tasks
- Prepare the solution for SDS-PAGE
- 500 mL Fixative Solution -> 40% methanol, 10% acetic acid, 50% water
- 500 mL Stain Solution -> 0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water
- 500 mL Destain Solution -> 10% acetic acid, 90% water)
- 200 mL of 10X SDS-PAGE Running Buffer -> 30.3 g Tris, 144.1g Clycine, 10g SDS, water to 1L + diluted x10 in 2000 mL of water
- non-dilute samples -> 10 µL reaction sample/10 µL running buffer. Enzyme concentration:
- Proteinase K Sample 1:
- 1 µM, 2 hrs
- 100 nM, 2.5 hrs
- 10 nM, 2 hrs
- 1 nM, 2 hrs
- Proteinase K Sample 2:
- 1 µM, modified protocol, 60 min
- 100 nM, modified, 80 min
- 10 nM, modified, 22 hrs
- 1 nM, modified, 21 hrs
- Thermolysin:
- 1 µM, 90 min
- 100 nM, 30 min
- 10 nM, 60 min
- 1 nM, 10 min
- Chymotrypsin:
- 1 µM, 1 hr
- 100 nM, 25 min
- 10 nM, 20 hrs
- 1 nM, 23 hrs
- Trypsin:
- 1 µM, 2 hrs
- 100 nM, 2.5 hrs
- 10 nM, 2 hrs
- 1 nM, 2 hrs
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