User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19: Difference between revisions
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#This morning at 8 am, the starter cultures were centrifuged at 4500 rpm for 15 minutes and then resuspended the cell pellets in 4mL of fresh LB. For more details on the process in the morning, see [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/09/19| Mary's Notebook]] | #This morning at 8 am, the starter cultures were centrifuged at 4500 rpm for 15 minutes and then resuspended the cell pellets in 4mL of fresh LB. For more details on the process in the morning, see [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/09/19| Mary's Notebook]] | ||
# Inoculate the [[AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media|expression culture media]] by dividing the resuspended cells among the Fernbach flasks. | # Inoculate the [[AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media|expression culture media]] by dividing the resuspended cells among the Fernbach flasks. | ||
--[[User:Abigail E. Miller|Abigail E. Miller]] 10:53, 7 October 2012 (EDT)it is not clear that this was done in the morning. | |||
# Incubate the expression cultures at 37°C and 160 rpm until the absorbance of the media at 600nm = 0.6. | # Incubate the expression cultures at 37°C and 160 rpm until the absorbance of the media at 600nm = 0.6. | ||
# Add 1mL of 0.4M [[AU_Biomaterials_Design_Lab:Materials/IPTG|IPTG]] to each flask in order to induce protein expression | # Add 1mL of 0.4M [[AU_Biomaterials_Design_Lab:Materials/IPTG|IPTG]] to each flask in order to induce protein expression | ||
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# Harvest the cells by centrifuging at 4500rpm for 15 minutes | # Harvest the cells by centrifuging at 4500rpm for 15 minutes | ||
# collect the cells and place in -80°C freezer | # collect the cells and place in -80°C freezer | ||
--[[User:Abigail E. Miller|Abigail E. Miller]] 10:53, 7 October 2012 (EDT)this is too much like the protocol. what did you actually do? at what time was the IPTG added? you did not measure the OD at 600nm. what time did you harvest the cells? therefore how long were they allowed to express? how many times did the cells have to be spun down? what deviations did you make from the protocol - I know you separated flask 1 form the other three. that is not clear here. | |||
==Calculations== | ==Calculations== |
Revision as of 07:53, 7 October 2012
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Protein Expression.Goals
Procedure
--Abigail E. Miller 10:53, 7 October 2012 (EDT)it is not clear that this was done in the morning.
--Abigail E. Miller 10:53, 7 October 2012 (EDT)this is too much like the protocol. what did you actually do? at what time was the IPTG added? you did not measure the OD at 600nm. what time did you harvest the cells? therefore how long were they allowed to express? how many times did the cells have to be spun down? what deviations did you make from the protocol - I know you separated flask 1 form the other three. that is not clear here. Calculations
a. Theoretical calculations
a. Theoretical calculations
Conclusions
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