User:Daniel-Mario Larco/Notebook/AU Photosynthesis Lab/2014/04/02: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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==Cell Extraction==
*The solutions from the flasks were centrifuged at 4700rpm for 15 minutes
*The pellet was collected and placed in a 50mL falcon tube with Tris buffer
*The solution was placed in a -80*C freezer until the next step


==Protein Expression==
#Cells were placed in each of the 25mL Erlenmeyer flasks and left to grow overnight in a shaking incubator at 37C and 200rpm
#Each of these solutions was transferred to a 1L broth solution with 1mL of ampicilin
#The flasks were left to incubate for 4 hours at 37C on the shaking incubator at 150rpm
#After 4 hours, 1ml of Fe-protoporphyrin IX was added to each flask and the temperature was reduced to 30C and left to incubate for another 4 hours
##The Fe-protoporphyrin IX was made by dissolving 40mg of Fe-protoporphyrin IX in 1mL of DMSO
#1mL of IPTG was also added to each flask
##4mM of IPTG in H20
#After four hours, the flasks were placed in a cold room over night
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Latest revision as of 23:53, 26 September 2017

Project name Main project page
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Protein Expression

  1. Cells were placed in each of the 25mL Erlenmeyer flasks and left to grow overnight in a shaking incubator at 37C and 200rpm
  2. Each of these solutions was transferred to a 1L broth solution with 1mL of ampicilin
  3. The flasks were left to incubate for 4 hours at 37C on the shaking incubator at 150rpm
  4. After 4 hours, 1ml of Fe-protoporphyrin IX was added to each flask and the temperature was reduced to 30C and left to incubate for another 4 hours
    1. The Fe-protoporphyrin IX was made by dissolving 40mg of Fe-protoporphyrin IX in 1mL of DMSO
  5. 1mL of IPTG was also added to each flask
    1. 4mM of IPTG in H20
  6. After four hours, the flasks were placed in a cold room over night


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