User:Jeffrey Kim/Notebook/Halogenated natural products from uncultured bacteria/2009/01/09

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TAR cloning progress

  • Clone ZB118 targeted with US and DS sequences (ABC transporter selected before NRPS system in ZB118).
  • PCR products indicate successful amplification with Buffer G at 65C for 30 cycles. (1, 2kb products see gel)
  • pLLX8 amplificaiton with TAR primers (ala Lory) look fine as well. Product is a bit dirty and yield was much lower so I repeated the experiment with a longer extension time (lanes 1 v 2).
  • EcoRI digest of the ten hits from recombinational cloning and assembly of ZB118 capture vector all look good. (released band of expected size, see gel)