AU Biomaterials Design Lab:Protocols/Transformation Protocol

Description

Transformation of PCR product into E. coli cells Transformation Manual

materials

Materials Supplied by the User

• Plasmid DNA (ready for transformation)
• 42°C water bath
• 37°C shaking and non-shaking incubator
• Ice bucket with ice
• Spectrophotometer to measure optical density of the cell cultures
• Microcentrifuge tube rack (optional)

Protocol

1. to PCR product, add 1 μL DPN1
2. place reaction at 37 °C for 1 hour

store in freezer till ready for trasnformation

1. prepare LB plates
2. Equilibrate water bath to 42 °C and place the LB plates at 37 °C

1. thaw onevial of cells on ice
2. add 10 ng of DNA in 5 μL to the cells ( DO NOT MIX BY PIPETTING)
3. heat shock the cells by incubating in 42 °C water bath for exactly 30 s
4. place immediately on ice