13/08/13

From 2013.igem.org

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.