IGEM:Cambridge/2008/Notebook/Voltage/2008/07/21

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Wet Work

Aims for Today

The results of yesterday's modified protocol still yielded no growth on the plates. Using yesterday's PCR product our aims were:

  • Cut promoters with spe1
  • Cut RBS with Xba1
  • Ligate these together and perform further PCR amplification of linear fragments.


  • Cut first 'stop' with spe1
  • Cut second 'stop with xba1
  • Ligate these together and perform further PCR amplification of linear fragments.


  • Finally, put all the above elements into a vector plasmid.


  • End of day: Aims completed

Length

  • Promoter = 35 bases
  • RBS = 16 bases
  • Stop (B1002) = 35 bases and Stop (B1006) = 39 bases