User:Matt Hartings/Notebook/Photosynthesis/2013/01/23
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(Autocreate 2013/01/23 Entry for User:Matt_Hartings/Notebook/Photosynthesis) |
Current revision (16:43, 23 January 2013) (view source) |
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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]] | |
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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. | ||
| + | |||
| + | ==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. | ||
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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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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) | |
