Jessica Karen Wong/Notebook/2007-7-26

Cleaning etc

• PCR cleaned all overnight digests - CCDB-E/X, CCDB-S/X, I2055-M/N, 3K3-E/P, 1AK3-E/P, 1AK3-E/X
• Minipreped 3K3 to have a DNA stock
• Made new Measurement Kit Primers box for the -20

I2055 Colony PCR Gel

I2055

• Ran gel of overnight col pcr's
• Only colony # 17 looks good
• Overnighted I2055-3K3 #17
• Ligated newly digested I2055 into the newly digested plasmids
  • Ligated I2055-3K3, I2055-1AK3
• Transformed 3ul ligation and plated

I2056

• Overnight transformation plate had no colonies
• Ligated I2056-1AK3, I2056-3K3 w/ newly digested plasmids
• Transformed and plated

E0240

BB PCR Gel

• Ligated E0240-1AK3
• Transformed and plated
• Set up 100ul BB PCR's of E0240-3K3 E/X and E0240-3K3 E/S at 52
• Ran gel of BB PCR, both look good (1st 2 bands after ladder)
• PCR cleaned both E0240-3K3-X and E0240-3K3-S
• Digested both BB PCRs w/ Not1 in buffer 3 overnight

I2057

• Ligated I2057-1AK3, transformed and plated
• BB PCRed I2057-3K3 E/X and I2057-3K3 E/S at 52
• Ran gel of BB PCR - both looked good (2 farthest right samples)
• PCR cleaned both BB pcr's
• Digest both w/ Not1 in buffer 3 overnight

T9002

• Overnight plates of T9002-1AK3 and T9002-3K3 have 4 colonies each
  • Set up 10ul Col PCR's
• Ligated F2620-1AK3 w/ new plasmid, transformed and plated
• Ran out of cut F2620
  • Digesting F2620 - M/X in buffer 4 overnight