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Team:Caltech/Week 14
From 2011.igem.org
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==September 13== | ==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/> | Quikchange of Wt-F87A with newly miniprepped DNA<br/> | ||
Plates of results, + the rest of SOC<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=== | ===Results=== | ||
No visible colonies on plates | No visible colonies on plates | ||
==September 15== | ==September 15== | ||
| - | + | Finish staining SDS gel<br/> | |
| - | SDS gel | + | |
Retransform quikchange pcr products<br/> | Retransform quikchange pcr products<br/> | ||
Make and autoclave M9 solution<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/> | 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|eluted DDT dechlorinase band clearly is visible, size according to broad range ladder</gallery><br/> | ||
==September 16== | ==September 16== | ||
Revision as of 17:57, 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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