User:Tk/Notebook/MF-xfm/2008/04/21

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Transformation results

  • E. coli transformation was normal 200+ colonies
  • picked six more colonies on replated osmolarity tests

recT gene PCR worked, expected size

  • Phusion in 100 ul final
    • 50 ul Phusion M/M
    • 1.5 ul recT-F
    • 1.5 ul recT-R primers
    • 1 ul 10 ng/ul S. citri genomic DNA (from Renaudin)
    • 46 ul water
  • cycle 98/30s (98/15s 55/15s 68/30s)x30 68/10 min
  • band visible at 900+ bp (expect 915)

Reporter design

  • most existing registry reporters have very bad codon usage
  • one possible exception is GFPmut3.1, E0040, which has pretty good codon usage but two Sau3AI sites