Transformation

Goal

In this step,

à modif

the heat shock (between -80°C up to 42°C) induces chimiocompetent bacteria to be in state of competence. DNA from the environment (in this case: plasmid) can then integrate the cell.

Preparation

Chimiocompetent cells

For 100 µL of chemical competent cells, add 1 µL of plasmidic DNA. If the ligation protocol is a Golden Gate , add 5 µL instead.
Let 30 minutes on ice.
Let the cells at 42°C for exactly 50 seconds.
Let 5 minutes on ice.
Resuspend the cells with 1 mL of LB spread on a petri dish then let it at 37°C.

Electrocompetent cells

Controles

To check if the transformation have work, make a positive and a negative controle.

Positive controle: Use plasmid pSB1A3
If the transformation have worked, colonies must have a red phenotype.

Negative controle: Ø
Without plasmid with antibiotic resistance, no colonies must grow.

Site developped by Gabriel Guillocheau