User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2016/01/14: Difference between revisions
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==Description== | ==Description== | ||
# Add | |||
Following Puja's methods | |||
# Heat a water bath to 80C | |||
# Degas water for making FeSO4 solution | |||
# Add the following to a falcon tube: | |||
## 1.87 mL of 1M KOH | |||
## 8.889 mL of 0.9M KNO3 | |||
## 2.338 mL of 15.4 uM Hemoglobin | |||
## 4.903 mL of water | |||
# Prepare the FeSO4 solution using the degassed water to prevent oxidation of Fe before it goes into the reaction | |||
## 2.085 g FeSO4 (MW 278.01 g/mol) | |||
## 25 mL degassed water | |||
# Place tube around mixing shaft | |||
# Place water bath around tube | |||
# Start mixing at 2500 rpm | |||
# Add 2 mL of FeSO4 solution | |||
# Let react for 90 minutes and place in freezer overnight | |||
Now I need to make a reaction that has no FeSO4 to look at the effects of just KOH and KNO3 | |||
# Heat a water bath to 80C | |||
# Add the following to a falcon tube: | |||
## 1.87 mL of 1M KOH | |||
## 8.889 mL of 0.9M KNO3 | |||
## 2.338 mL of 15.4 uM Hemoglobin | |||
## 6.903 mL of water | |||
# Place tube around mixing shaft | |||
# Place water bath around tube | |||
# Start mixing at 2500 rpm | |||
# Add 2 mL of FeSO4 solution | |||
# Let react for 90 minutes and place in freezer overnight | |||
I also need to prep solutions to run electrophoresis | |||
# 10X running buffer | |||
## 30.3 g Tris | |||
## 144.1 g glycine | |||
## 10 g SDS | |||
## Add water to 1L | |||
## stir on stir plate overnight to mix solution | |||
# 2X sample buffer | |||
## 3.75 mL of 0.5 M Tris (pH 6.8) | |||
## 7.5 mL of glycerol | |||
## 0.3 mL of 1% bromophenol blue | |||
## 0.6 g SDS | |||
## H2O to 30 mL | |||
==Data== | ==Data== |
Revision as of 11:21, 14 January 2016
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ObjectiveThe referees of the microgel paper wanted me to determine if the protein hydrolyzes during the reaction for making hydrogels. So, I'm going to run 2 reaction (one with iron and one without iron) to see if the protein is degraded by the KOH and KNO3. DescriptionFollowing Puja's methods
Now I need to make a reaction that has no FeSO4 to look at the effects of just KOH and KNO3
I also need to prep solutions to run electrophoresis
Data
NotesThis area is for any observations or conclusions that you would like to note.
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