Jessica Karen Wong/Notebook/2007-8-1

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I2057

  • Transformation plates w/ DB3.1 had no colonies
  • Doing a positive control on DB3.1 w/ puc18
  • Was going to digest the preps of I2057-EX and I2057-ES resealed w/ Not1, but didn't have enough DNA
  • Making a 10ml overnight culture of each
  • Minipreped overnight cultures of promoters J23116 and J23100
  • Digested R0040, J23116, and J23100 - Eco/Spe

I2056

  • PCR cleaned overnight digests of I2056-EX w/ E/X, I2056-EX w/ Not, I2056-SX S/X, I2056-SX w/ Not1
  • Ligated I2056-EX-N and I2056-SX-N each to themselves (closed up)
  • Transformed in Top10 and plated
  • Digested RBS's B0030, B0031, and B0032 E/S and X/S each for later testing

I2055

BB I2055 PCR
  • Ran a gel of the overnight BB PCR's loaded: ladder sp I2055-EX sp I2055-SX
  • Looked the right size
  • PCR Cleaned overnight PCRs
  • Digesting I2055-SX with S/X, I2055-SX w/ Not1, I2055-EX with E/X, I2055-EX w/ Not1 overnight

F2620

  • Transformation plates had no colonies
  • Religated F2620-3K3
  • Transformed with Top10 and plated
  • We're running out of DNA, so made new PCR
  • Ran 1 100ul supermix PCR to add tails to F2620 in 1A3 at 53 w/ 1:30 ext time