Week2 6/20/10-6/26/10

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Week2 Summary

We began making antibiotic plates. We also took out parts 12O and 6G from the iGEM kit because we planned on testing inducible promoters. We ran into our first problem when we received low DNA yields after DNA extraction. We also experienced weird reddish/pink colonies in our 12O plates which we expected to be completely white.


We poured our Kan, Amp, Tet, Kan+Amp, Kan+Tet, Amp+Tet antibiotic agar plates.

Antibiotic Concentrations

  • Amp: 50mg/500ml
  • Kan: 31.97mg/500ml
  • Tet: 6mg/500ml


Resuspended 12O and 6G DNA from the iGEM Kit and transformed them into our competent cells.

  • 6G = r0011(inducible promoter)
  • 12O = e0840(rbs30-gfp-2xterm)


Ran a gel of our PCR product to make sure that our taq polymerase master mix was working.

Selected single 12O and 6G colonies and transferred them to LB and incubated for 18 hours.

Let 1 12O and 1 6G plate incubate at RT. Let 1 12O and 1 6G plate incubate at 37°C. Intended to observe lawn growth.


Low DNA yield; Plan to redo DNA extraction from Kit

Found red/pink colonies in 12O plates. Thought to be contaminations. Plates were thrown out.