IGEM:IMPERIAL/2006/LabCalendar/2006-7-21

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T9002 Test Assay


  • Part T9002 and R0062 taken out of incubation at 1100.
  • OD Reading:
    • T9002 = 1.792
    • R0063 = 1.857
  • The cultures were diluted 10x and incubated from 1130-1350
  • OD Reading:
    • T9002 = 0.610
    • R0063 = 1.150
  • Cultures were diluted 2x and incubated from 1430-1500
  • OD Reading:
    • T9002 = 0.682
    • R0063 = 1.048

The rest of the experiment followed the protocol


Digest

  • Made digest of the parts minipreped yesterday and ran gels A-D
  • Gel 'A' was defective (marker and part failed to be 'sufficiently' contained in the well...we reckon it still worked though...)
  • Most of the parts worked although some were not compatible for testing on the same gel (parts of small molecular weight just ran off the gel and so couldn't be seen. May require re-testing for assurance.)
  • Results:
  1. Gels A-C
  2. Gels C-D

For all the gels the far left well contained the molecular marker. We labelled the gels A-D; each well is labelled 1-6 (1 being the molecular marker)

Gel A:

  • well 1A: Molecular marker
  • well 6A: C0160 (perhaps successful - probably needs retesting)

Gel B:

  • well 1B: Molecular marker
  • well 2B: E0040 (successful)
  • well 3B: F1610 (successful)
  • well 4B: C0260 (successful)
  • well 5B: B0034 (part very small - ran off gel)
  • well 6B: E0434 (successful)

Gel C:

  • well 1C: Molecular marker
  • well 2C: I0403 (successful)
  • well 3C: F2620 (successful)
  • well 4C: C0261 (successful)
  • well 5C: R0062 (part very small - ran off gel)
  • well 6C: T9002 (successful)

Gel D:

  • well 1D: Molecular marker
  • well 2D: I14032 (part very small - ran off gel)
  • well 3D: I13207 (successful)
  • well 4D: C0062 (failed to identify part)
  • well 5D: I0460 (no plasmid band)
  • well 6D: I13504 (successful)


We think some of the 'extra' bands may be due to supercoiling. Some of the thicker bands could be due to insufficient digestion of DNA.

C0062 and I0460 definitely need retesting, as they have unexplainable bands. R0062, I14032 and B0034 have very small parts (55, 37 and 12 base-pairs respectively) and so require retesting, making sure that they are only left to run for a short period of time. C0160 may possibly have been successful, but as the gel that it was on was defective, it is probably best to retest this part as well.

We will retest these parts on Wednesday 26th July, when we digest some other new parts that we are growing up.

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