Team:Uppsala-SwedenWeek14

From 2010.igem.org

Construction Of K1, K2, K3

The inoclum obtained from the overnight culture was used for plasmid extraction. The extracted plasmids were cut at specific restriction sites present in the biobrick sequence and run on the gel.

The gel was checked for correct fragment lengths to perform a second round f validation The extracted plasmids were digested with the correct set of enzymes as defined in the flow chart.

The extracted plasmids were purified and used for performing ligation with the correct prefix or suffix and the appropriate vector. The ligate was used to perform transformation of competent cells The competent cells were plated on appropriate cells for transformation.

The concentration of plasmid were measured for all the samples.

Plasmid concentration of all starting biobricks.Plasmid of J1, J2, J3 were digested with proper enzymes. The biobricks were ligated in pairs according to the plan by Quick Ligase kit from Fermentas.

The ligation product were transformed into DH5α competent cells and each sample were plated on the agar plate which antibiotics matched to its backbone antibiotic resistance.