User:Matt Hartings/Notebook/Photosynthesis/2012/09/12: Difference between revisions
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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 08:36, 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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