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Lab Work - 11/09/09

Metal Sensing Team

Introduction and Outline

So far we have transformed DH5-alpha E. coli cells with two of our synthesized BioBricks, which are the cotC-GFP-smtA and the kinA BioBricks. The Metal Sensing team have since taken these transformant colonies and used them to set up LB+kanamycin cultures for mini-preps and midi-preps; these were then left overnight in the orbital shaking incubator and are awaiting processing today. However there are a range of tasks which need to be accomplished today; these are listed below:

• PCR the cotC-GFP-smtA BioBrick with pMK15 and pMK16
  • Set up the PCR
  • Run the PCR for the correct duration
  • Analyse PCR products through DNA gel electrophoresis
  • Clean up PCR products if PCR reaction proves to be successful
• Midi-prep cotC-GFP-smtA, kinA, pGFP-rrnB, pMUTIN4 and pSB1AT3
  • Carry out the five midi-preps.
  • Quantify the midi-prep samples
  • Run the midi-prep samples on agarose gel through DNA gel electrophoresis
• Run products from a PCR reaction involving arsR and cadA DNA
  • Run PCR products on gel
  • If successful, clean up fragments, run on gel again, cut with restriction enzymes NheI and BamHI, clean and then ligate together fragments.