Registry/Measurement kit/Notebook/2007-7-3

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Sequence verification

  • Notes
    • Use AlignX for multiple sequence alignment
    • Sequences in Documents -> sequences -> folder named w/date
    • View chromatograms in FinchTV
    • Registry also has sequence verification software in development.
  • I2055 sequence didn't check (R0040 missing)
    • This explains why the primers for the PCR didn't work...
  • Choosing colonies 3,4, and 6 to sequence, since they looked slightly larger on the 6/19 gel.
    • Growing with amp+kan
  • Also ordered primer to insert the sequence by PCR:
    • format: 8-mer.MfeI.promoter+mix.part of gfp (Tm)
    • CTTAGTAG.CAATTG.tccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactag.ATGCGTAAAGGAGAAGAACTTTTC (52.4)

I2056 Construction

  • Got one colony on 1AT3
  • Re-plating the rest (1AT3, 1AC3)
  • Did colony PCR, but reaction seems to have evaporated...diluting w/10uL water and running on gel. re-doing PCR in parallel

Analytical Gel

  • Loaded in this order: ladder, blank, t9002 scarred, I2056, ladder.
Gel from 7/3/07

2 log ladder