Jessica Karen Wong/Notebook/2007-7-27

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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


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