Team:Panama/20 July 2010

From 2010.igem.org

(Difference between revisions)
(July 20)
(Activities:BioBrick extraction from the distribution plates)
 
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'''Methodology Steps:'''
'''Methodology Steps:'''
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'''1.Locate the BioBrick parts that we are going to assemble, in the distribution plates.'''
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'''1.Locate the BioBrick parts that we are going to assemble in the distribution plates.'''
RBS: 2M, GFP: 14K, Terminator: 13D & Promoter: 15N
RBS: 2M, GFP: 14K, Terminator: 13D & Promoter: 15N
-
For that we take the DNA from each of the wells of the plate. We then have to take 2ul of each one and put it in the bacteria. The other 8ul we are to store at -20°C.
 
-
'''2.Clone the chosen BioBrick parts in competent ''E. coli''.'''  
+
'''2. Resuspend the plasmid part with 10ul water (Nuclease free, distilled)'''  
-
For the extraction we follow the comercial protocol. We transformed 4 different bacteria, for the different BioBrick plasmids & prepared LB-ampi. [[Image:Cuadro_medio1.JPG|200px|thumb|left|alt text]] [[Image:Plates2.JPG|400px|thumb|left|alt text]]
+
For that we take the DNA from each of the wells of the plate. We used 2ul of each part to performed the E. coli transformation. The remaining 8ul were store at -20°C.
 +
 
 +
 
 +
'''3.Transformed the ''E. coli'' with the chosen BioBrick parts.'''
 +
 
 +
For the extraction we follow the comercial protocol. We transformed 4 different bacteria, for the different BioBrick plasmids.  
 +
 
 +
 
 +
[[Image:Cuadro_medio1.JPG|200px|thumb|left|alt text]] [[Image:Plates2.JPG|400px|thumb|left|alt text]]
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For the ampicilin we have to make a frozen stock solution with a concentration of 50ug/ml in water. In the plate we need a concentration of 100mg/ml.
+
For the ampicilin we made a frozen stock solution with a concentration of 50ug/ml in water. In the plate we used a concentration of 100mg/ml.
-
We want to make a stock solution of 2ml so, we need 100mg of ampicilin to make 50mg/ml. We have to filter this through a 0.2um filter.
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Latest revision as of 03:22, 27 October 2010

iGEM Panama

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July 20

Activities:BioBrick extraction from the distribution plates

Methodology Steps:

1.Locate the BioBrick parts that we are going to assemble in the distribution plates.

RBS: 2M, GFP: 14K, Terminator: 13D & Promoter: 15N


2. Resuspend the plasmid part with 10ul water (Nuclease free, distilled)

For that we take the DNA from each of the wells of the plate. We used 2ul of each part to performed the E. coli transformation. The remaining 8ul were store at -20°C.


3.Transformed the E. coli with the chosen BioBrick parts.

For the extraction we follow the comercial protocol. We transformed 4 different bacteria, for the different BioBrick plasmids.


alt text
alt text


Incubate at 37°C overnight LB agar plates were prepared with Ampicillin, pH 7.5



For the ampicilin we made a frozen stock solution with a concentration of 50ug/ml in water. In the plate we used a concentration of 100mg/ml.


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