Recombinant DNA Lab

          The first step of producing recombinant DNA is to splice open the plasmid. Our plasmid had bacteria that was resistant to ampicillin. We could use ampicillin in our petri dishes to see if bacteria had taken over the plasmid since we had the gene that was resistant to it. We would know if the bacteria had taken our plasmid because if it did then the bacteria would be resistant to the antibiotic and survive. I wouldn't use any of the other antibiotics because our bacteria isn't resistant to any of the others and it would have just killed all the bacteria. After that we tested all of the restriction enzymes to see which ones could splice open the plasmid in one place and the human DNA in two places. Restriction enzymes are bacterial enzymes that recognize a specific nucleotide sequence in DNA molecules and cut the molecules at that sequence. Two of the ones we tested spliced the plasmid and human DNA the correct amount of times. The two enzymes were Eco RI and Hin dIII; however, although both of them worked, the Hin dIII spliced closer to the insulin gene. Therefore, we ended up using the Hin dIII. If we had used an enzyme that cut the plasmid in two places then it would not have joined together with the DNA and it wouldn't have been able to make recombinant DNA. There would have been two extra sticky ends. We then found the areas where the bases of the restriction enzyme and the bases of the human DNA and plasmid had been spliced. The sticky ends then joined with the DNA pieces and formed recombinant DNA. This technology could be important in daily life because it allows the insertion of different genes to give an organism new traits that could possibly prevent/cure certain diseases. A real life example of this technology being used is in the creation of genetically engineered plants that produce a toxin called Bt. It kills off certain crop pests so that farmers' crops don't get eaten away at by insects and have higher success rates.