User:Matt Hartings/Notebook/Photosynthesis/2012/09/12: Difference between revisions
From OpenWetWare
(Autocreate 2012/09/12 Entry for User:Matt_Hartings/Notebook/Photosynthesis) |
(fix raw html notebook nav) |
||
(2 intermediate revisions by one other user not shown) | |||
Line 1: | Line 1: | ||
{|{{table}} width="800" | {|{{table}} width="800" | ||
|- | |- | ||
|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> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
Line 11: | Line 11: | ||
==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) | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Latest revision as of 21:59, 26 September 2017
Protein Re-engineering | Main project page Previous entry Next entry |
ObjectiveRun WT Hb expression with non iron protoporphyrin IX to see if expression proceeds as normal Procedure
|