13 August 2013

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Making agar, chloroamphenicol and chloroamphenicol/IPTG plates

  • One container of 400 ml Luria Agar was melted down in the microwave. This is enough to make 20 agar petri dishes.
  • This was poured straight into 4 petri dishes to make agar plates
  • The remaining agar had 0.64 ul of chloroamphenicol added to it (concentration = 24 ug/ml), and was mixed thoroughly
  • The resulting solution was poured into 4 petri dishes, to make 4 chloroamphenicol agar plates
  • The remaining agar solution had 1.5 ul of IPTG added to it (concentration = 24 ug/ml)
  • This was mixed thoroughly, and then used to make up the remaining 12 chloroamphenicol/IPTG plates.

Agarose gel preparation

  • 20 ml of 5 x TBE, 80 ml of distilled water and 0.8 g of agarose was added into a container.
  • The solution was boiled for 3 minutes, 1 minute at a time.
  • The solution was left to cool.
  • 5 ul of ethidium bromide was added to the solution.
  • The solution was then poured into the gel plate and left to set for 30 minutes.