Exeter/18 July 2013

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Exeter iGEM 2013 · Paint by Coli

NanoDrop data from yesterday's gel extraction

The NanoDrop machine was zero-ed using the elution buffer from the gel extraction kit used.

  • Yellow pigment (BBa_K592010) - 6.6ng/µl
  • Magenta pigment (BBa_K592012) - 4.9ng/µl
  • Green light sensor, CcaS (BBa_K592001) - 3.2ng/µl
  • RFP control - 4.4ng/µl
  • New Part 2, promoter and RBS added to FixJ - 2.1ng/µl
  • Blue light sensor, YF1 (BBa_K592004) - 1.4ng/µl
  • FixJ protein intermediate, (BBa_K592005) - 10.0ng/µl
  • Kanamycin resistant plasmid - 2.0ng/µl

These concentrations are reasonably low, so the SureClean protocol from 17/7/13 was implemented.

Part First concentration (ng/µl) New concentration (ng/µl)
Yellow pigment 6.6 168.7
Magenta pigment 4.9 154.6
CcaS (green light sensor) 3.2 81.1
"New Part 2" (promoter, RBS and FixJ) 2.1 207.6
YF1 (blue light sensor) 1.4 172.8
FixJ 10.0 155.8

We also concentrated the amounts of DNA in five replicates of our terminator (BBa_B0015).

Part Concentration (ng/µl)
Terminator #1 200.8
Terminator #2 193.6
Terminator #3 183.0
Terminator #4 182.6
Terminator #7 190.9

Ligations

Name Contents
New Part 1 CcaS (green light sensor) + terminator
New Part 2 (promoter, RBS and FixJ) + terminator
New Part 3 Yellow pigment + terminator
New Part 4 Magenta pigment + terminator
New Part 7 YF1 (blue light sensor) + terminator
New Part 8 FixJ + terminator

Re-streak plating

We re-streak plated all of the plates that were more than 10 days old, which were:

  • Magenta pigment (K592012)
  • OmpR promoter
  • RBS (B0034) PSB1A2
  • Terminator (B0015) PSB1L3
  • Blue sensor (K592004) PSB1C3
  • Promoter + RBS (K608002) PSB1C3
  • CcaS (K592001)
  • RFP control
  • FixJ intermediate (K592005)
  • CcaR
  • GFP
  • Yellow pigment (K592010)


  • NOTE : The GFP and yellow pigment plates got muddled up before they were labelled and had to be streaked the next day (19th July).
  • NOTE : When the RFP control had been incubated over night it had turned a magenta/red colour.

Take me back to the notebook.

Exeter iGEM 2013 · Paint by Coli