Team:Caltech/Week 3

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Revision as of 06:50, 29 June 2011

Caltech iGEM 2011

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Data

Parts

Team

Notebook

Biosafety

Human Impact

References

Support

June 26

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])

June 27

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
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)

Results

Miniprep

Part	Concentration(ng/ul)
B0014	230.5
B0015	254.1
J06702	301.3
K145001	205.6
R0010	117.3
R0040	156.8

We found growth in one of our BPA enrichment cultures, and slight growth in our ethinyl estradiol cultures. We transferred these vials today.

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

Results

PCR of parts for pNT001 and pNT002: From left to right: 1 ladder; 2 blank; 3 [http://partsregistry.org/Part:BBa_R0010 R0010]; 4 [http://partsregistry.org/Part:BBa_K123000 K123000]; 5 [http://partsregistry.org/Part:BBa_B0014 B0014]; 6 [http://partsregistry.org/Part:BBa_R0040 R0040]; 7 [http://partsregistry.org/Part:BBa_K123001 K123001]; 8 [http://partsregistry.org/Part:BBa_B0015 B0015]

Ran a gel of the PCR products. Strangely, lanes 3 and 4 show no DNA. We will repeat the PCR of lanes 3-5 and Gibson assemble pNT001 ([http://partsregistry.org/Part:BBa_R0010 R0010], [http://partsregistry.org/Part:BBa_K123000 K123000], [http://partsregistry.org/Part:BBa_B0014 B0014]) later in the week.

EtOH precipitation of Soil Extractions from June 21

Tube/Location Number	Concentration(ng/ul)
1	13.6
2	15.2
4	8.3
5	11.4
6	25.5
7	23.5
9	261.3
10	74.0

This Week

PCR of parts for pNT001 (Test plasmid for [http://partsregistry.org/Part:BBa_K123000 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 55: / Line 55: @@
 Gibson assembly of pNT001 and pNT002</p>
 ===Results===
+[[File:628pcrgel.png|thumb|PCR of parts for pNT001 and pNT002: From left to right: 1 ladder; 2 blank; 3 [http://partsregistry.org/Part:BBa_R0010 R0010]; 4 [http://partsregistry.org/Part:BBa_K123000 K123000]; 5 [http://partsregistry.org/Part:BBa_B0014 B0014]; 6 [http://partsregistry.org/Part:BBa_R0040 R0040]; 7 [http://partsregistry.org/Part:BBa_K123001 K123001]; 8 [http://partsregistry.org/Part:BBa_B0015 B0015] ]]
+Ran a gel of the PCR products. Strangely, lanes 3 and 4 show no DNA. We will repeat the PCR of lanes 3-5 and Gibson assemble pNT001 ([http://partsregistry.org/Part:BBa_R0010 R0010], [http://partsregistry.org/Part:BBa_K123000 K123000], [http://partsregistry.org/Part:BBa_B0014 B0014]) later in the week.<br/><br/>
 EtOH precipitation of Soil Extractions from [[Team:Caltech/Week 2|June 21]]<br/>
 <table border="1">
@@ Line 94: / Line 96: @@
 </tr>
 </table>
 == This Week ==
 <p>PCR of parts for pNT001 (Test plasmid for [http://partsregistry.org/Part:BBa_K123000 K123000])<br/>