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From 2010.igem.org

Week-13

Identification of biobricks

Plasmid of the samples which showed right size band on the gel were sent to a plasmid concentration test. We decided only J1, J2, J3 were in good concentration to proceed.

However, we got the correct PCR lengths for Bio-bricks and results are shown in image files below. .The result was showed in Fig.1 and Fig2

                             J1-J3

                             J2-J3

biobricks

However, we got the correct band lengths for all J1, J2, J3. And the one that are with correct band lengths were selected. The correct length samples of J1, J2, J3 were again inoculated again to get the enough plasmid for further progress.

Second colony PCR was performed again for all the starting biobricks and the products were run for gel again. Comparing the DNA length to the band size, all the biobricks were confirmed.

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 bought from Sigma-Aldrich. 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.