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Culture Parts for Testing

We will be testing the zinc hypothesis by using the part MIT used in 2004 (I13207). This part as well as T9002 was cultured in 2 mL LB Amp overnight for testing tomorrow. We are thinking of using 4 different tests which will tell us if having zinc in the medium makes all the difference.

  • M9, no Zn, with Arabinose (Control)
  • M9, with Zn, with Arabinose
  • LB, with Arabinose
  • LB, no Arabinose (Control)

M9 medium protocol can be found on OpenWetWare.

  • Also note that to make the Amp LB for Maxi-prep (Midi-prep), go down to level one of Biochem building to the Media Office to order. Bring 500 mL Erlynmeyer flasks and ask for: 100 mL LB with 50 ug/mL Amp.

  • Ligations and part 1I have been minipreped
  • Gel is made ready to be run tomorrow

To do on Tuesday 1 August:

  • 30 uL digests of both 7A and 3O
  • Run large gel to extract DNA fragments
    • For 7A, we want the 756 bp one, NOT 2000 bp fragment
    • For 3O, we want the 2091 bp one, NOT the small fragment
    • Set gel at 130 V, 0.130A to run gel
    • Extract fragments using transilluminator
  • Ligate parts overnight