20.109(S11): TA notes for module 2

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20.109(S11): Laboratory Fundamentals of Biological Engineering

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General notes

Key preparation:


Scheme:

Day-by-day

Day 1

NOT YET COMPLETE - SEE ALSO GOOGLE DOC

Materials required:

  • Two days before lab, streak out the following strains:
    • ABX(8?) = pEDL3/pCph8/pPLPCB, on Amp/Cam/Kan plate
    • ABX(22?) = pED-IPTG-INS, on Amp plate
  • One day before lab, prepare O/N cultures of same
    • AB8? is for edge detection plates
    • AB22? is for first liquid culture experiment

Below are at set up at teaching bench:

  • 2 boxes of cuvettes

Day of Lab (T/R):

  • Prepare supplemented LB medium (30' autoclave, 30' or 60' for pressure to go down in large or small autoclave, respectively) and cool in a 42 °C water bath for at least 1 hour

After Lab

How it went:

Day 2

Etc.

Special materials

Cell strains

  • NB399 = JW3367c (EnvZ-, lacZ-, Kan^R removed)
  • NB435/AB? = pEDL3, pCph8, pPL-PCB (full ED system)
  • NB? = light to lacZ only
  • AB? = pED-IPTG-110
  • AB? = pED-IPTG-112
  • AB? = pJT104 (full ED system less AHL)
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