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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.
82 run on plate reader during day
85 overnight
Wednesday
Wet Work
Threshold Devices
Confirmed successful ligation of pBad and I746350, I746351, I746352.
90 run during day
92 run overnight
Thursday
Wet Work
Threshold Devices
94 run during day
95 overnight
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 as a PoPS converter (below).
The next step will be to attach the phage promoter downstream to create a catalogue of 15 different devices of the form:
Friday
Wet Work
Threshold Devices
Running a test to see if 40 was successfully moved into pSB3K3
75 run during day
91 overnight
