Wednesday, January 20, 2010
Today we used the purified PCR product from the nested PCR lab (GAPC gene from Arabidopsis) to genetically transform E. coli.
The lab was divided in three main steps
- Preparation of competent cells
- Ligation
- Genetic transformation
During most of the lab students manipulated 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 that were spread on LB agar/Amp/IPTG plates and incubated.
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