Jessica Karen Wong/Notebook/2007-8-10

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  • Minipreped overnights
  • Sent for seq - I2056-ENX (3,5,7,8) I2056-SNX (1,5,6,8) I2055-SX-B34 (2 colonies)
  • Transformation plates looked better (after leaving them to rest for a day)
  • Made overnights of I2055-EX-B34, I2055-SX-B23, I2056-SX-B32, I2056-EX-B34, I2056-SX-B34 (2 colonies of each)
    • Also made an overnight of the streaked plate of I2057-EX-J116 (to separate the 2 types), probably will still be mixed
F2620 Colony PCR Gel

  • Set up colony PCRs of the 3 plates of F2620 in DB3.1
  • Ran a gel loaded plate 1 (8-1) sp lad sp plate 2 (8-1), w/ plate 3 (8-1) on 2nd row
  • Made overnights of f2620 #2 colonies 6 and 3, F2620 #3 colonies 3 and 4

  • Ligated:
    • I2056-B0032 EX PCR
    • I2056-B0031 EX and SX PCR
    • I2055-B0032 EX and SX Prep
    • I2055-B0031 EX and SX prep
  • Trying to put B0031 and B0032 in all variations of RBS testers
  • Set up a Dpn1 digest of all I2055 and I2056 ligations (just added to the ligation mix)
  • Also set up another BB PCR with 1ng of DNA
    • Made all versions I2055-EX, I2055-SX, I2056-EX, I2056-SX


  • I2055-SX-B30 - both samples (glowing and not glowing) had each a 1bp mutation in gfp
  • E0240-ES-M2 bad read, ES site is there and 1st 100bp look good, should redo w/ gfp internal primer
  • I2055-EX both failed!
  • I2055-SX both completely have part but contain no BB site
  • I2055-EX-B0032 nowhere near, not sure what that was
  • I2056-7 scar, not necessary already know it was scarred
  • I2057-ES looks good
  • I2057-EX BB site is there, runs till 344, if you compare w/ 1st attempt to seq you get full coverage!