The products of restriction digestion were separated

The products of restriction digestion were separated on 1.0% agarose gel that had been stained with µg/ml 0.5ethidium bromide. First, two samples consisting of the pBKS II plasmid digest and Pc-FtsZ insert were placed on the gel containing 5µl loading buffer. The nest step involved running the gel in a tank containing TAE buffer. A MW marker was loaded into the first and last wells on the gel. 20µl of each sample was loaded into different wells. Similarly, undigested samples were loaded into the gel to act as controls. The gel was run at 110V for 50 min to separate the fragments into distinct bands.

The bands corresponding to the plasmid and insert pattern were cut out before being transferred into a micro-centrifuge tube. Its mass was determined before adding QG buffer (three volumes). Subsequent steps involved repeated incubation, dissolution in isopropanol, and centrifugation of the samples at maximum speed to bind the samples to the spin column. The spin column was then washed in EB buffer to elute the DNA. The purified DNA was stored at -20°C for the next step.