Difference between revisions of "User:Matt Hartings/Notebook/Photosynthesis/2013/01/23"

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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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==Minipreps==
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From yesterday.
  
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Only the swMb cells grew. I'll have to try the pQE-80 again in amp just to check ...
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I extracted the DNA using the Wizard miniprep [http://www.promega.com/~/media/Files/Resources/ProtCards/Wizard%20Plus%20SV%20Minipreps%20DNA%20Purification%20System%20Quick%20Protocol.pdf protocol] and reagents.
  
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I measured the DNA concentrations for the 4 samples prepared using UV Vis (diluting each sample 2uL sample into 200uL total).
  
==Objective==
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[[Image:WTswMbMiniprep 20130123.png|300px]]
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The final concentrations were estimated to be: 30, 40, 25, and 35 ug/mL respectively.
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I combined all of the samples, placed them in an eppy tube, poked holes in the cap, froze the eppy/sample, and lyophilized them for 5 hours. After all of the solvent had evaporated, I labeled the sample pT7-7/WT swMb lyophilized, and placed it in the freezer in the myoglobin box.
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==Lyophilizing==
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I removed the samples from yesterday's Mb lyophilizing. However, there was still solid water in the samples. So I returned them to the lyophilizer.
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==MOF==
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Abigail removed the MOF synthesis (from yesterday) from the oven at around 9:30am to allow the sample to cool down. There was a white ppt at the bottom of the flask and the surrounding liquid was yellow. Abigail filtered the sample. We still need to analyze the solid (by powder x-ray diffraction) and the liquid (by UV-Vis, Fluorescence, mass spec, LC)
  
 
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Latest revision as of 21:23, 26 September 2017

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

Minipreps

From yesterday.

Only the swMb cells grew. I'll have to try the pQE-80 again in amp just to check ... I extracted the DNA using the Wizard miniprep protocol and reagents.

I measured the DNA concentrations for the 4 samples prepared using UV Vis (diluting each sample 2uL sample into 200uL total).

WTswMbMiniprep 20130123.png

The final concentrations were estimated to be: 30, 40, 25, and 35 ug/mL respectively.

I combined all of the samples, placed them in an eppy tube, poked holes in the cap, froze the eppy/sample, and lyophilized them for 5 hours. After all of the solvent had evaporated, I labeled the sample pT7-7/WT swMb lyophilized, and placed it in the freezer in the myoglobin box.

Lyophilizing

I removed the samples from yesterday's Mb lyophilizing. However, there was still solid water in the samples. So I returned them to the lyophilizer.


MOF

Abigail removed the MOF synthesis (from yesterday) from the oven at around 9:30am to allow the sample to cool down. There was a white ppt at the bottom of the flask and the surrounding liquid was yellow. Abigail filtered the sample. We still need to analyze the solid (by powder x-ray diffraction) and the liquid (by UV-Vis, Fluorescence, mass spec, LC)