Lab Notebook

27 July 2009

Calin

Placed GFP / RFP plates in fridge. Ordered some trimethoprim. Placed pSB1AK3 cells in tube for culturing. Finished writing Matlab code to plot parameter space. Cleared up confusing equations for mRNA and protein.

Sriram

Prepared 1 litre of LB-Agar and separated in 4x500 ml beakers and sent for autoclaving with a beaker of eppendorfs. Since it was not received before 18:00 just diluted the 6 plasmid backbone biobricks that will be used in this project. Looked into the protocol that will be used for transformation of them in ccdB resistant competent cells.

Ran the agarose gel electrophoresis with the 10 biobricks [R0040, J23008, J23031, B0015, K081013, S03335, S03473, I714031, E0840, J23100] for 1 hour to check the extracted DNA. It must be continued tomorrow hence the gel was preserved in fridge.

Inoculated the top10 cells for preparing competent cells and along with it the two colonies got from

Orr

Used mfold to produce 3 locks: one with weak RBS, one with weaker RBS and one with strong RBS.
Using the 2-state hybridization server, combined the key3c with each of the 4 locks (lock3c, weak lock, weaker lock and strong lock) to obtain the hybridised structure.
Created weak key, weaker key and strong key sequences for each of the last three locks by taking the hybridization of each lock with the key3c and correcting the kinks by changing the key3c sequence.
Biobricked the weak, weaker and strong keys into the registry.

Tim Weenink

Nanodropped the plasmid isolations I did on friday:
The *I is the code for the biobrick I have constructed, that contains the I-SceI cleavage site. The !A is an abbreviation for a brick containing RBS32 + GFP-LVA.

I also did some restrictions (all in µl) (incubated for 35min at 37ºC, then inactivated):

And some more (10min 37ºC, 20min inactivation):

Then loaded the gel: