Team:Caltech/Week 14

From 2011.igem.org

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__NOTOC__
__NOTOC__
==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==
-
 
+
Quikchange of Wt-F87A with newly miniprepped DNA<br/>
-
 
+
Plates of results, + the rest of SOC<br/>
==September 14==
==September 14==
-
 
+
Overexpression of his-tagged DDT-dechlorinase in pET protein expression vector in preparation for purification, at 20&degC and 37&degC<br/>
 +
===Results===
 +
No visible colonies on plates
==September 15==
==September 15==
-
Protein purification <br/>  
+
Protein purification of DDT-dechlorinase using Sigma-aldrich his-select protein purification columns<br/>
-
Stretch gel <br/>  
+
SDS gel of purified products, as well as pellet, first flow-through, and first wash<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/>
==September 16==
==September 16==
-
Desalting DDT gene enzyme with 2L 20mM Tris-HCI & 150 mM NaCI <br/>  
+
Made desalting buffer (20mM Tris-HCl + 150mM NaCl)<br/>
-
 
+
Desalt DDT-dechlorinase buffer of imidazole to stabilize protein through dialysis<br/>
-
==September 17==
+
Made X-gal-IPTG plates for quikchange lacZ repair control<br/>
-
<br/>
 
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Revision as of 07:09, 19 September 2011


Caltech iGEM 2011



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September 12

Miniprep of WT-F87A

Results

Miniprep concentrations:

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

September 13

Quikchange of Wt-F87A with newly miniprepped DNA
Plates of results, + the rest of SOC

September 14

Overexpression of his-tagged DDT-dechlorinase in pET protein expression vector in preparation for purification, at 20&degC and 37&degC

Results

No visible colonies on plates

September 15

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

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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