Jessica Karen Wong/Notebook/2007-7-27

Colony PCRs

• Ran gel of F2620-3K3 and F2620-1AK3 overnight colony pcr's
  • Saw nothing, redoing the colony pcr's
• Overnight plates mostly looked good
  • I2057-1AK3 plate had 7 colonies
  • I2055-1AK3 had 5
  • I2056-1AK3 had 1
  • I2055-3K3 and I2056-3K3 had lots
  • E0240 had none but looked dried out
  • F2620-3K3 and F2620-1AK3 had none/way too small ones
• Spun down and replated E0240-1AK3, I2057-1AK3, I2055-1AK3, I2056-1AK3, F2620-1AK3
• Ran Colony PCR's of I2057-1AK3 (7 colonies), I2055-1AK3 (5 col), I2056-1AK3 (1 col), I2056-3K3 (8 col), I2055-3K3 (8 col)
• Ran overnight colony PCR's of 16 more I2055-3K3

Digests and Ligations

• Cleaned overnight digests: E0240-X Not1, E0240-S Not1, F2620 M/X, I2057-X Not, I2057-S Not
• Digested: E0240-X E/X, E0240-S E/S, I2057-X E/X, I2057-S E/S, CCDB in 1AT3 E/S
• Ligated: F2620-3K3, F2620-1AK3, E0240-X to close, E0240-S to close, I2057-X to close, I2057-S to close
• Transformed and plated each ligation
• Made new KAN plates

Gels

I2057, I2056, I2055, F2620 Colony PCR Gel

• Ran gels of today's Colony PCR's

I2055 and I2056 Colony PCR Gel

• Large Gel Top Row: I2057-1AK3 (1-7 and blank) sp lad sp I2056-3K3 (1-8)
• Large Gel Bottom Row: I2055-3K3 (1-8) sp lad sp F2620-3K3 (3-1) F2620-1AK3 (4-1) F2620-3K3
• Small Gel: lad sp I2055-1AK3 (5-1) I2056-1AK3
• Overnighted I2055-3K3 colony #1 and I2056-3K3 Colonies 1 and 2