Talk:BE.109:Protein engineering/Purifying beta-galactosidase
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To compare the purification protocols you have tried today, each team in each lab section should enter their data into the following tables.
Tuesday/Thursday
Team Color | purification method | equation for BSA standard curve | concentration of beta-gal stock | concentration of purified beta-gal (wild type) | activity of purified beta-gal (wild type) | concentration of modified beta-gal (list modification below table) | activity of modified beta-gal |
---|---|---|---|---|---|---|---|
Green | Ni-agarose | y=.7133x - .0244 | 2.08 mg/ml | .29 mg/ml | 27595 units | .158 mg/ml | 1822 units |
Purple | Paramagnetic Beads | OD-595 = 0.5556(Conc.) - 0.0012 | 4.19 mg/mL | 0.0501 mg/mL | 37.37 units | 0.0506 mg/mL | 34.87 units |
Red | Ni-agarose | y=.7893x-0.0274 | 2.345 mg/mL | 0.874 mg/mL | .076 | .56 mg/mL | .085 |
Blue! | Ni-agarose | 0.6709x-0.0002 | 3.75mg/mL | 0.647 | 0.0318 | 0.322 | 0.233 |
Pink | Ni-agarose | y=1.2x+0.0002 | 2.332 mg/mL | 0.111 mg/mL | OD 420= 0.7363 , Units = 2.6783 | 0.268 mg/mL | OD 420= 0.2173, Units= 1.8339 |
Yellow | paramagnetic beads | y = 1.11x-.06 | .55 mg/mL | .50 mg/mL | OD420=.56, Units = 48890.86 | .61 mg/mL | OD420=.63, Units = 34878.04 |
Wednesday/Friday
Team Color | purification method | equation for BSA standard curve | concentration of beta-gal stock | concentration of purified beta-gal (wild type) | activity of purified beta-gal (wild type) | concentration of modified beta-gal (list modification below table) | activity of modified beta-gal |
---|---|---|---|---|---|---|---|
Green | Ni-agarose | y = 0.6258x - 0.0197 | 3.39 mg/mL | 0.473 mg/mL | 0.0356 umol/min/mg | 0.543 mg/mL (modified His) | 0.0362 umol/min/mg |
Purple | |||||||
Red | |||||||
Blue | |||||||
Pink | Ni-agarose | y=.9833x-.0906 | 1.1899 mg/mL | 0.2801 mg/mL | 3.8357*10^8 umol/min/mg | 0.3363 mg/mL (modified Glu) | 5.2358*10^8 umol/min/mg |
Yellow |