Team:Caltech/Week 3

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Revision as of 00:35, 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
Check DDT and nonylphenol cultures, transfer to fresh minimal media if there is growth
Ethanol precipitation of DNA extractions from LA river samples
Streak out cells from fosmid kit
Fosmid kit first gel?
Prepare antibiotic stocks (Ampicillin, Chloramphenicol)

June 28

Send off continued forward sequencing for HER
Observe fosmid gel, conduct shearing and end repair, run size selection gel (if needed)
Start culture of fosmid cells overnight

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

@@ Line 7: / Line 7: @@
 <p> Start overnight cultures 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])</p>
-== This Week ==
+==June 27==
-<p>Transfer 0.5 mL aliquots of BPA and 5 mL aliquots 17a-ethynylestradiol cultures from June 23 to fresh minimal media<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/>
+Check DDT and nonylphenol cultures, transfer to fresh minimal media if there is growth <br/>
 Ethanol precipitation of DNA extractions from LA river samples<br/>
-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/>
+Streak out cells from fosmid kit<br/>
-PCR of parts for pNT001 (Test plasmid for [http://partsregistry.org/Part:BBa_K123000 K123000])<br/>
+Fosmid kit first gel? <br/>
+Prepare antibiotic stocks (Ampicillin, Chloramphenicol)</p>
+==June 28==
+<p>Send off continued forward sequencing for HER <br>
+Observe fosmid gel, conduct shearing and end repair, run size selection gel (if needed)<br/>
+Start culture of fosmid cells overnight </p>
+== This Week ==
+<p>PCR of parts for pNT001 (Test plasmid for [http://partsregistry.org/Part:BBa_K123000 K123000])<br/>
 Purification/Gibson assembly for pNT001<br/>
-Cutting LA River DNA for insertion into fosmids<br/>
 Email other teams <br/>
-Insert LA River DNA into fosmids <br/>
-Transform fosmids into E. coli and start enrichment cultures<br/>
-Continue gene fishing <br/>
-Send off continued forward sequencing for HER<br/>
 PCR of parts for pNT002 and assembly<br/>
 Use pNT001 and pNT002 to test degradation parts</p>
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