Team:Caltech/Week 3

From 2011.igem.org

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<p>Miniprep of T7 polymerase ([http://partsregistry.org/Part:BBa_K145001 K145001]), mCherry ([http://partsregistry.org/Part:BBa_J06702 J06702]), Lac Promoter ([http://partsregistry.org/Part:BBa_R0010 R0010]), double terminator ([http://partsregistry.org/Part:BBa_B0014 B0014] and [http://partsregistry.org/Part:BBa_B0015 B0015]) and Tet Promoter ([http://partsregistry.org/Part:BBa_R0040 R0040]) and submit for sequencing<br/>
<p>Miniprep of T7 polymerase ([http://partsregistry.org/Part:BBa_K145001 K145001]), mCherry ([http://partsregistry.org/Part:BBa_J06702 J06702]), Lac Promoter ([http://partsregistry.org/Part:BBa_R0010 R0010]), double terminator ([http://partsregistry.org/Part:BBa_B0014 B0014] and [http://partsregistry.org/Part:BBa_B0015 B0015]) and Tet Promoter ([http://partsregistry.org/Part:BBa_R0040 R0040]) and submit for sequencing<br/>
Transfer 0.5 mL aliquots of BPA and 5 mL aliquots 17a-ethynylestradiol cultures from June 23 to fresh minimal media<br/>
Transfer 0.5 mL aliquots of BPA and 5 mL aliquots 17a-ethynylestradiol cultures from June 23 to fresh minimal media<br/>
-
Check DDT and nonylphenol cultures, transfer to fresh minimal media if there is growth <br/>
+
Checked DDT and nonylphenol cultures, will wait until tomorrow to transfer<br/>
-
Ethanol precipitation of DNA extractions from LA river samples<br/>
+
Ethanol precipitation of DNA extractions from LA river samples following protocol<br/>
Streak out cells from fosmid kit<br/>
Streak out cells from fosmid kit<br/>
-
Fosmid kit first gel? <br/>
 
Prepare antibiotic stocks (Ampicillin, Chloramphenicol)</p>
Prepare antibiotic stocks (Ampicillin, Chloramphenicol)</p>
==June 28==
==June 28==
<p>Send off continued forward sequencing for HER <br>
<p>Send off continued forward sequencing for HER <br>
-
Observe fosmid gel, conduct shearing and end repair, run size selection gel (if needed)<br/>
+
Prepare gel of control fosmid DNA to practice gel extraction technique<br/>
-
Start culture of fosmid cells overnight </p>
+
PCR for Gibson<br/>
-
 
+
Gibson assembly of pNT001 and pNT002</p>
== This Week ==
== This Week ==

Revision as of 00:38, 28 June 2011


Caltech iGEM 2011



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June 26

Start overnight cultures of T7 polymerase (K145001), mCherry (J06702), Lac Promoter (R0010), double terminator (B0014 and B0015) and Tet Promoter (R0040)

June 27

Miniprep of T7 polymerase (K145001), mCherry (J06702), Lac Promoter (R0010), double terminator (B0014 and B0015) and Tet Promoter (R0040) and submit for sequencing
Transfer 0.5 mL aliquots of BPA and 5 mL aliquots 17a-ethynylestradiol cultures from June 23 to fresh minimal media
Checked DDT and nonylphenol cultures, will wait until tomorrow to transfer
Ethanol precipitation of DNA extractions from LA river samples following protocol
Streak out cells from fosmid kit
Prepare antibiotic stocks (Ampicillin, Chloramphenicol)

June 28

Send off continued forward sequencing for HER
Prepare gel of control fosmid DNA to practice gel extraction technique
PCR for Gibson
Gibson assembly of pNT001 and pNT002

This Week

PCR of parts for pNT001 (Test plasmid for K123000)
Purification/Gibson assembly for pNT001
Email other teams
PCR of parts for pNT002 and assembly
Use pNT001 and pNT002 to test degradation parts


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