Jessica Karen Wong/Notebook/2007-7-24

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I2056 Scar/Promoter Gel


  • Ran a gel of the overnight PCR
  • Looks good
  • Digesting with Mfe and Nsi in buffer 2 overnight


T9002 E0240 Gel
  • Got new T9002_E0240_R primer in - diluted to 40 uM
  • Set up a 100ul supermix PCR rxn of E0240 for T9002 to put on tails
    • Ran at 52
  • Ran a gel which looks great
  • PCR cleaned
  • Digesting w/ Spe/Nsi in buffer 2 overnight


  • Made a new overnight from the registry plate of 1AK3
  • Got sequencing back for I2055-3K3 - really wierd
    • 1st half of part (till middle of GFP) was put in backwards, rest of part was forwards
  • Found that I2055 has an Mfe1 site in the middle of GFP and that's where the problem happened
  • Changed synthesis proposal so doesn't have the Mfe1 site in GFP - silent mutation