Jessica Karen Wong/Notebook/2007-7-26

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Contents

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 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
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
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