Team:TUDelft/Temperature results

From 2008.igem.org

(Difference between revisions)
(Results)
(Results)
Line 8: Line 8:
[[Image:TUDelftschematic3A.png|thumb|400px|center|Figure 1. Representation of the 3A assembly method. The black arrow indicates the [http://partsregistry.org/Part:BBa_P1010 CcdB 'death' gene]. The arrows in other colors indicate different types of antibiotic resistance. Restriction sites are indicated by: E = ''Eco''R1; P = ''Pst''I; X = ''Xba''I; S = ''Spe''I]]
[[Image:TUDelftschematic3A.png|thumb|400px|center|Figure 1. Representation of the 3A assembly method. The black arrow indicates the [http://partsregistry.org/Part:BBa_P1010 CcdB 'death' gene]. The arrows in other colors indicate different types of antibiotic resistance. Restriction sites are indicated by: E = ''Eco''R1; P = ''Pst''I; X = ''Xba''I; S = ''Spe''I]]
 +
 +
<br/>
 +
For the succesful use of this assembly, it is essential that the DB3.1 ''Eschirichia coli'' strain is not used as it tolerates the presence of the CcdB gene. Throughout the project we used commercial Top10 cells of Invitrogen that were made competent chemically. Furthermore, it is important to note that the ''Xba''I and ''Spe''I restriction enzymes generate compatible sticky ends.
{{Template:TUDelftiGEM2008_sidebar}}
{{Template:TUDelftiGEM2008_sidebar}}

Revision as of 09:41, 28 October 2008

Results

The first challenge of this project was to assemble all the devices we wanted to measure. An overview of these devices can be found here. For the actual construction of all devices the 3A assembly strategy was chosen. A complete protocol of this strategy can be found on [http://openwetware.org/wiki/Synthetic_Biology:BioBricks/3A_assembly OpenWetWare]. A schematic representation of this assembly method is depicted in figure 1.

Figure 1. Representation of the 3A assembly method. The black arrow indicates the [http://partsregistry.org/Part:BBa_P1010 CcdB 'death' gene]. The arrows in other colors indicate different types of antibiotic resistance. Restriction sites are indicated by: E = EcoR1; P = PstI; X = XbaI; S = SpeI


For the succesful use of this assembly, it is essential that the DB3.1 Eschirichia coli strain is not used as it tolerates the presence of the CcdB gene. Throughout the project we used commercial Top10 cells of Invitrogen that were made competent chemically. Furthermore, it is important to note that the XbaI and SpeI restriction enzymes generate compatible sticky ends.