Team:Caltech/Week 8

From 2011.igem.org

(Difference between revisions)
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Transformation and plating of pUC19 into competent cells for cell dry weight experiment<br/>
Transformation and plating of pUC19 into competent cells for cell dry weight experiment<br/>
Overnight culture of B0034 glycerol stock<br/>
Overnight culture of B0034 glycerol stock<br/>
 +
Overnight ligation of pNT002 insert and PSB3K3 backbone<br/>
 +
Restriction digest of parts for pNT002 standard assembly and overnight ligation <br/>
===Results===
===Results===
Line 37: Line 39:
Gibson assemble pNT003 using the gel extracted insert and linearized pSB4A5<br/>
Gibson assemble pNT003 using the gel extracted insert and linearized pSB4A5<br/>
Replating of more pUC19-transformed competent cells and non-transformed competent cells<br/>
Replating of more pUC19-transformed competent cells and non-transformed competent cells<br/>
 +
Run gel of overnight pNT002 ligations<br/>
 +
Transformed pNT002 insert+ pSB3K3 ligation<br/>
===Results===
===Results===
<gallery>
<gallery>
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File:8-2pcrgel2.jpg|lane 1 NEB 2-log ladder, 2-4 pNT003 insert, 5-7 pNT003 inset with PIPE primers, 8-9 pNT003 insert, 10-11 pNT003 inset with PIPE primers, 12-15 pETDEST53
File:8-2pcrgel2.jpg|lane 1 NEB 2-log ladder, 2-4 pNT003 insert, 5-7 pNT003 inset with PIPE primers, 8-9 pNT003 insert, 10-11 pNT003 inset with PIPE primers, 12-15 pETDEST53
</gallery>
</gallery>
 +
pNT002 ligation showed band on gel
}}
}}

Revision as of 00:15, 3 August 2011


Caltech iGEM 2011



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July 31

Started a LB+amp overnight culture of the B0034 glycerol stock from the 2010 Caltech iGEM Team

Results

Mass spectrometry did not run correctly; samples did not ionize; must rerun

August 1

PCR pSB3K3 and pNT003 insert with the new primers.
Gel extract the pNT002 insert from the July 29 PCR
Minimal media transfers
Transformation and plating of pUC19 into competent cells for cell dry weight experiment
Overnight culture of B0034 glycerol stock
Overnight ligation of pNT002 insert and PSB3K3 backbone
Restriction digest of parts for pNT002 standard assembly and overnight ligation

Results

No colonies visible on the pUC19-transformed cells

The enrichment culture plates show no visible growth yet.

Gel Extractions of July 29 PCR of pNT002 and pNT003 inserts

Part Concentration (ng/ul)
pNT002 insert 65.9
pNt003 insert 12.5

August 2

Gibson assemble pNT002 using the gel extracted insert and linearized pSB3K3
Gibson assemble pNT003 using the gel extracted insert and linearized pSB4A5
Replating of more pUC19-transformed competent cells and non-transformed competent cells
Run gel of overnight pNT002 ligations
Transformed pNT002 insert+ pSB3K3 ligation

Results

lane 1 NEB 2-log ladder, 2-4 pNT003 insert, 5-7 pNT003 insert with PIPE primers, 8-13 pSB3K3, 14-15 pETDEST53

lane 1 NEB 2-log ladder, 2-4 pNT003 insert, 5-7 pNT003 inset with PIPE primers, 8-9 pNT003 insert, 10-11 pNT003 inset with PIPE primers, 12-15 pETDEST53

pNT002 ligation showed band on gel


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