User:Matt Hartings/Notebook/Photosynthesis/2012/09/12: Difference between revisions
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|style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> Protein Re-engineering</span> | ||
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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 08:50, 12 September 2012
Protein Re-engineering | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ObjectiveRun WT Hb expression with non iron protoporphyrin IX to see if expression proceeds as normal Procedure
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