User:Matt Hartings/Notebook/Photosynthesis/2013/01/23: Difference between revisions
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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"| | |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}}}}]]}} | ||
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==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 [http://www.promega.com/~/media/Files/Resources/ProtCards/Wizard%20Plus%20SV%20Minipreps%20DNA%20Purification%20System%20Quick%20Protocol.pdf protocol] and reagents. | |||
I measured the DNA concentrations for the 4 samples prepared using UV Vis (diluting each sample 2uL sample into 200uL total). | |||
[[Image:WTswMbMiniprep 20130123.png|300px]] | |||
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) | |||
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Latest revision as of 22:23, 26 September 2017
 Protein Re-engineering
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MiniprepsFrom 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).
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. LyophilizingI removed the samples from yesterday's Mb lyophilizing. However, there was still solid water in the samples. So I returned them to the lyophilizer.
MOFAbigail 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) | |
