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Team:Caltech/Week 14
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__NOTOC__ | __NOTOC__ | ||
==September 12== | ==September 12== | ||
| - | + | Miniprep of WT-F87A | |
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===Results=== | ===Results=== | ||
Miniprep concentrations: | Miniprep concentrations: | ||
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</table><br/> | </table><br/> | ||
| + | ==September 13== | ||
| + | Overexpression of his-tagged DDT-dechlorinase in pET protein expression vector in preparation for purification, at 20°C and 37°C<br/> | ||
| + | Quikchange of Wt-F87A with newly miniprepped DNA<br/> | ||
| + | Plates of results, + the rest of SOC<br/> | ||
==September 14== | ==September 14== | ||
| - | + | Protein purification of DDT-dechlorinase using Sigma-aldrich his-select protein purification columns<br/> | |
| + | SDS gradient gel of purified products, as well as pellet, first flow-through, and first wash<br/> | ||
| + | ===Results=== | ||
| + | No visible colonies on plates | ||
==September 15== | ==September 15== | ||
| - | + | Finish staining SDS gel<br/> | |
| + | Retransform quikchange pcr products<br/> | ||
| + | Make and autoclave M9 solution<br/> | ||
| + | Set up biofilm experiments using different substrates (glass beads vs. wood chips), media (dilute LB vs. M9), and conditions (20°C vs. 37° C vs. shaking)<br/> | ||
| + | ===Results=== | ||
| + | <gallery>File:DDTdechlorinase.jpg|Purified DDT dechlorinase band visible, size according to BioRad broad range protein marker</gallery><br/> | ||
==September 16== | ==September 16== | ||
| - | + | Made desalting buffer (20mM Tris-HCl + 150mM NaCl)<br/> | |
| - | + | Desalt DDT-dechlorinase buffer of imidazole to stabilize protein through dialysis<br/> | |
| - | + | Made X-gal-IPTG plates for quikchange lacZ repair control<br/> | |
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Latest revision as of 18:16, 19 September 2011
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Project |
September 12Miniprep of WT-F87A ResultsMiniprep concentrations:
September 13Overexpression of his-tagged DDT-dechlorinase in pET protein expression vector in preparation for purification, at 20°C and 37°C September 14Protein purification of DDT-dechlorinase using Sigma-aldrich his-select protein purification columns ResultsNo visible colonies on plates September 15Finish staining SDS gel Results
September 16Made desalting buffer (20mM Tris-HCl + 150mM NaCl)
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