Team:Cambridge/Notebook/Week9

From 2009.igem.org


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.


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

Wet Work

Threshold Devices

Running a test to see if 40 was successfully moved into pSB3K3

75 run during day

91 overnight

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