Module 2, Lab 06 - Ligation and transformation (GAPC gene from Arabidopsis and pJet1.2 plasmid)

Today we purified the PCR products from the nested PCR lab (GAPC gene from Arabidopsis) and used them to genetically transform E. coli.

The lab was divided in three main steps

• Ligation (of GAPC gene on to the pJet1.2 plasmid)
• Preparation of competent cells
• Genetic transformation of E. coli

We spent most of the lab manipulating bacteria to make them competent (i.e. get them ready to uptake extracellular DNA). Once this was achieved, the GAPC gene from Arabidopsis, obtained via nested PCR, was ligated to the pJet1.2 plasmid.
The plasmid was then used to genetically transform E. coli, which were spread on LB agar/Amp/IPTG plates and incubated.

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