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April 27, 2009

  1. Received from Rosa (SPiT):
    • TM0785
      • Plasmid containing encapsulin
      • Recommend transfect into bacteria and re-sequence
      • See email note regarding sequence error
    • 0.5 microliters TMG DNA 100 microgram/microliter
      • Use 0.4 microliter for 50 microliter PCR reaction =August 1, 2009=
  2. Overnight encapsulin cultures 1 and 6 (randomly selected) and the negative control were miniprepped
  3. The rest of the encapsulin cultures were stocked with 20% glycerol
  4. 6 digestions were done: Enc1, Enc6, Enc -ve, BB5, BB7, and C
    • 16ul of BB5 plasmid was used
    • 500ng of plasmid were used for the others
  5. The digestions were run on a 1.3% agarose gel in TAE (from here on, unless otherwise specified, all gels were 1.3% agarose)
    • BB5 was confirmed and all other parts were correct as well
  6. Overnight ligation of 7+Enc in the PCR machine