Difference between revisions of "User:Matt Hartings/Notebook/Photosynthesis/2013/04/04"

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|style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> Protein Re-engineering</span>
 
|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"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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==Objective==
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==Mb catalysis DMSO==
Learn how to maintain an OpenWetWare Notebook.
+
Following the description from [[User:Matt_Hartings/Notebook/Photosynthesis/2013/04/02|Tuesday]].
 +
<u>Stock solutions used</u>
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# 18.4mg of o-phenylenediamine in 17mL of DMSO
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# 1.134 ml 30% H2O2 +8.866 ml DMSO
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# 11.1mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1 mL of DMSO.
  
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<u>Samples were made by</u>
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# 2.025 mL of the OPD solution
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# 0.225 mL of the Mb solution
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# Start the scan
 +
## 1,000 us integration time
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## Average over 500 scans
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## Run for 10 minutes (600,000,000 us)
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# Add 0.75mL of the H2O2 solution
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## Added after scan reached 1%
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# Kinetics scans were run at 25, 35, and 45C (DMSO freezes at around 20C). Several before and after spectra were also taken, as were the dark and reference signal spectra.
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# Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]
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 +
==Mb catalysis H2O==
 +
Following the description from [[User:Matt_Hartings/Notebook/Photosynthesis/2013/04/02|Tuesday]].
 +
# 22.0mg of o-phenylenediamine in 22mL of H2O
 +
# 1.134 ml 30% H2O2 +8.866 ml H2O
 +
# 15.0mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1.5 mL of H2O.
 +
 +
<u>Samples were made by</u>
 +
# 2.025 mL of the OPD solution
 +
# 0.225 mL of the Mb solution
 +
# Start the scan
 +
## 1,000 us integration time
 +
## Average over 500 scans
 +
## Run for 10 minutes (600,000,000 us)
 +
# Add 0.75mL of the H2O2 solution
 +
## Added after scan reached 1%
 +
# Kinetics scans were run at 5, 15, 25, 35, and 45C. Several before and after spectra were also taken, as were the dark and reference signal spectra.
 +
# Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]
  
 
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Latest revision as of 21:36, 26 September 2017

Hartings AU Photosynthesis Lab Header.png Protein Re-engineering Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png



Mb catalysis DMSO

Following the description from Tuesday. Stock solutions used

  1. 18.4mg of o-phenylenediamine in 17mL of DMSO
  2. 1.134 ml 30% H2O2 +8.866 ml DMSO
  3. 11.1mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1 mL of DMSO.

Samples were made by

  1. 2.025 mL of the OPD solution
  2. 0.225 mL of the Mb solution
  3. Start the scan
    1. 1,000 us integration time
    2. Average over 500 scans
    3. Run for 10 minutes (600,000,000 us)
  4. Add 0.75mL of the H2O2 solution
    1. Added after scan reached 1%
  5. Kinetics scans were run at 25, 35, and 45C (DMSO freezes at around 20C). Several before and after spectra were also taken, as were the dark and reference signal spectra.
  6. Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]

Mb catalysis H2O

Following the description from Tuesday.

  1. 22.0mg of o-phenylenediamine in 22mL of H2O
  2. 1.134 ml 30% H2O2 +8.866 ml H2O
  3. 15.0mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1.5 mL of H2O.

Samples were made by

  1. 2.025 mL of the OPD solution
  2. 0.225 mL of the Mb solution
  3. Start the scan
    1. 1,000 us integration time
    2. Average over 500 scans
    3. Run for 10 minutes (600,000,000 us)
  4. Add 0.75mL of the H2O2 solution
    1. Added after scan reached 1%
  5. Kinetics scans were run at 5, 15, 25, 35, and 45C. Several before and after spectra were also taken, as were the dark and reference signal spectra.
  6. Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]