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Team:Cambridge/Notebook/Week9
From 2009.igem.org
Categories :
Project :
-
Overview
Sensitivity Tuner
--- Characterisation
--- Modelling
Colour Generators
--- Carotenoids (Orange/Red)
--- Melanin (Brown)
--- Violacein (Purple/Green)
The Future
Safety
Notebook :
Team Logistics :
Week 9
Monday
Wet Work
Threshold Devices
Overnight cultures of 70, 71, 72, 74, 91 in pSB3K3 in arabinose strain in order to make glycerol stocks and streak single colonies for the plate reader.
Tuesday
Wet Work
Threshold Devices
All activator constructs are now ready for analysis on the plate reader.
Wednesday
Wet Work
Threshold Devices
Confirmed successful ligation of pBad and I746350, I746351, I746352.
Thursday
Wet Work
Threshold Devices
Attempted the following standard assemblies:
- pBad + I746350 to B0015
- pBad + I746351 to B0015
- pBad + I746352 to B0015
- I746351 to B0015
- I746352 to B0015
- I746352 to B0015
The first three will be used to construct a complete device, with pBad as the sensor promoter and a pigment operon as the pigment-generating device. The pigment we chose for our proof of concept will be placed downstream of each of the 5 phage promoters, and then combined to give 15 combinations of activators and promoters, giving the construction below. We still need to decide which pigment to use for our proof of concept.
The second three will be used to construct a library of threshold devices which can be abstracted to black boxes with PoPS in and PoPS out (below). The next step will be to attack the phage promoter downstream to create a catalogue of 15 different devices.
Friday
