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DNA and Restriction Enzyme Interaction Identification Of An Unknown Plasmid Abstract The fundamental purpose of these experiments is to research the interaction between DNA molecules and restriction enzymes Two different experiments are performed In the first experiment two restriction enzymes Hind III and Bam H1 interact with an unknown plasmid and the resulting DNA fragments are separated through electrophoresis Lambda sample is used to create a standard curve and through it the sizes of DNA fragments are determined An uncut sample is used as a negative control and is compared with cut samples to verify the enzyme activities The size of the unknown sample is determined to be 4200 base pairs through the single cut sample In the double cut sample the sizes of fragments are determined to be 2350 and 1850 which concludes that the unknown sample is pKAN In the transformation experiment TE pAMP pKAN and two unknowns are put in five different tubes E-coli is used as the host bacterium The TE serves as a negative control because is does not contain DNA The pAMP and pKAN serve as positive controls and colonize in ampicilin and kanamycin media respectively The orange-labeled unknown shows identical phenotype with pKAN thus its genotype confirmed to be pKAN The green-labeled unknown shows identical phenotype with pAMP thus its genotype confirmed to be pAMP Introduction Modern genetic engineering began in 1973 when Herbert Boyer and Stanley Cohen used enzymes to cut a bacteria plasmid - ring of extra DNA found outside the nucleus in many single-celled organisms the relaxed control plasmid was used in experiments because it produced a lot more copies of plasmid than stringent control plasmid1 - and insert another strand of DNA in the gap2 Both bits of DNA were from the same type of bacteria but this milestone the invention of recombinant DNA technology offered a window into the previously impossible -- the mixing of traits between totally dissimilar organisms
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