Team:Caltech/Week 14

From 2011.igem.org

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__NOTOC__
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==September 12==
==September 12==
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Miniprep of WT-F87A
===Results===
===Results===
Miniprep concentrations:
Miniprep concentrations:
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</tr>
</tr>
</table><br/>
</table><br/>
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==September 13==
==September 13==
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+
Overexpression of his-tagged DDT-dechlorinase in pET protein expression vector in preparation for purification, at 20&degC and 37&degC<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==
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+
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&degC vs. 37&deg 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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==September 17==
 
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Latest revision as of 18:16, 19 September 2011


Caltech iGEM 2011



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References

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September 12

Miniprep of WT-F87A

Results

Miniprep concentrations:

Miniprep Nanodrop conc. (ng/uL)
WT-F87A 118

September 13

Overexpression of his-tagged DDT-dechlorinase in pET protein expression vector in preparation for purification, at 20&degC and 37&degC
Quikchange of Wt-F87A with newly miniprepped DNA
Plates of results, + the rest of SOC

September 14

Protein purification of DDT-dechlorinase using Sigma-aldrich his-select protein purification columns
SDS gradient gel of purified products, as well as pellet, first flow-through, and first wash

Results

No visible colonies on plates

September 15

Finish staining SDS gel
Retransform quikchange pcr products
Make and autoclave M9 solution
Set up biofilm experiments using different substrates (glass beads vs. wood chips), media (dilute LB vs. M9), and conditions (20&degC vs. 37&deg C vs. shaking)

Results


September 16

Made desalting buffer (20mM Tris-HCl + 150mM NaCl)
Desalt DDT-dechlorinase buffer of imidazole to stabilize protein through dialysis
Made X-gal-IPTG plates for quikchange lacZ repair control


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