Team:Cambridge/Notebook/Week9

From 2009.igem.org

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All activator constructs are now ready for analysis on the plate reader.  
All activator constructs are now ready for analysis on the plate reader.  
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90 run during day
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92 run overnight
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Revision as of 14:21, 11 September 2009


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.

82 run on plate reader during day 85 overnight

Tuesday

Wet Work

Threshold Devices

All activator constructs are now ready for analysis on the plate reader.

90 run during day

92 run overnight

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.


Proofofconcept.jpg


The second three will be used to construct a library of threshold devices which can be abstracted as a PoPS converter (below).

Thresholddeviceabstraction.jpg

The next step will be to attach the phage promoter downstream to create a catalogue of 15 different devices of the form:

Completedevice.jpg


Friday

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