User:Matt Hartings/Notebook/Photosynthesis/2012/09/12
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==Objective== | ==Objective== | ||
| - | + | Run WT Hb expression with non iron protoporphyrin IX to see if expression proceeds as normal | |
| + | ==Procedure== | ||
| + | #Spin down starter cultures from the previous evening. | ||
| + | #Resuspend in 4mL fresh media | ||
| + | #Add 1mL of 100mg/ml ampicilin to each 2.8L fernbach flask | ||
| + | #Divide the 4mL resuspended cells among each expression flask | ||
| + | #Place flasks in shaker at 37C | ||
| + | #Grow to an OD of 0.6 at 600nm (Note: the cells grew to an OD of 0.9) | ||
| + | #Reduce temperature to 30C | ||
| + | #Add 1mL of 0.4M IPTG to each flask | ||
| + | #Add 1mL of metal-protoporphyrinIX to each flask | ||
| + | ##40mg of Mn-protoporphyrin IX in 1mL of DMSO | ||
| + | ##40mg of Ni-protoporphyrin IX in 1mL of DMSO (Note: I only had 16mg of this complex) | ||
| + | ##40mg of Co-protoporphyrin IX in 1mL of DMSO | ||
| + | ##40mg of Cu-protoporphyrin IX in 1mL of DMSO | ||
| + | #Shake for 4 hours after induction (I induced at 11:00am) | ||
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Revision as of 11:36, 12 September 2012
 Biomaterials Design Lab
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ObjectiveRun WT Hb expression with non iron protoporphyrin IX to see if expression proceeds as normal Procedure
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