Contact In the news Acknowledgements References Links Downloads Protocols Brainstorm Meetings

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 OpenWetWare. A schematic representation of this assembly method is depicted in figure 1A and 1B.

Figure 1A. Representation of the 3A assembly method. The black arrow indicates the 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

Figure 1B. The mixture present in the ligation mix before it is ligated during 3A assembly

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.