Team:NYU/Clipboard

From 2010.igem.org

Today I transformed biobricks necessary for the second half of our system construction. Over the weekend I've been brainstorming ways to construct the response system circuits and have decided that we'll be using the LexA and LacI DNA binding proteins. The Initial Distribution this year came with most of the parts we'll need including some very useful composite reporter devices. For a time I also organized our growing collection of liquid and glycerol biobricks to maintain smaller stocks, something that will probably be an ongoing battle in the war against disorganization.

We are having some trouble assembling very small parts onto larger ones, case in point: the RBS onto a large promoter. Harris believes that the dichotomy of sizes of these parts makes it hard to relatively represent each in reaction solutions. We thought that the PCR product assembly method would work for this but have been disappointed thus far. For the sake of time I am going to order oligos and PCR-ligate the RBS onto pGAL and pTEF, as well as the RBS onto LacO onto pTEF for our reporter constructs. Once this is finished we'll hopefully be able to work smoothly through the assembly procedure. At this point I would probably give up a kidney for the ability to synthesize all of these constructs.

Also sent ECFP:Terminator I and II to sequencing to be 100% certain they are of the correct sequence.