Jessica Karen Wong/Notebook/2007-8-7

I2056 BB PCR Gel

• Ran a gel of the overnight I2056 BB pcr's
  • Loaded: Lad EX SX
  • Both look right size
• Heat Shocked I2055 S/X digest from PCR
• Minipreped E0240-EX, E0240-ES, F2620 for seq
• PCR cleaned I2056 pcr's and I2055-SX digest
• Digested - E2056-EX both E/X and Not1, I2056-SX both S/X and Not1
• Sent for seq each w/ VF and VR
  • F2620 to make sure it's in correct
  • E0240-EX colonies E2, E1
  • E0240-ES colonies M2, M1, E2
  • I2057-EX-J100 (1, 2)
  • I2057-ES-R40 (Bright, 1, 2)
  • I2057-EX-R40 (1,2)
  • I2057-EX-J116 (1,2)
  • I2055-SX-B31
• Most I2055 plates had very few fluorescent protein
  • We realized that we didn't use restriction enzymes in ligation mix
  • Were going to do again w/ adding restriction enzymes to ligation mix, but didn't have enought DNA
  • Overnighted I2055-SNX and I2055-ENX
• Overnighted 2 colonies of I2055-SX-B0030, 1 that fluoresced and 1 that didn't