%0 Journal Article
%T Cloning DNA through the use of Recombinant DNA Technology
%A Erik Rotenhoffen (MSc)
%J School of Doctoral Studies Journal
%D 2010
%I 
%X Recombinant DNA technology was used to clone specific DNA fragments. Before the technology was used, a basic understanding of the parts and details of the procedure were investigated. It was found that the larger the genome of an organism, the greater its overall complexity relative to other living organisms. This was discovered through the comparison of various genome sizes coming from known specimen. Gel electrophoresis technology was utilized to compare the relative sizes of the DNA. The gels were run from left to right with increasing level of complexity. The samples were, from left to right, plasmid DNA, lambda bacteriophage DNA, calf thymus DNA, and calf kidney DNA. As the complexity of the DNA increased, the number of bands in the lane increased to the point that the Eukaryotic DNA found in the calf thymus and kidney were smears. The gel displayed identical smearing and satellites for the calf kidney and thymus DNA, proving that the DNA is the same even though they are from different part of the organism. Through DNA fingerprinting, DNA fragments were determined that matched a specific pattern after being exposed to a restriction enzyme (HindIII). The DNA fragment containing the 2.0 ¦Ë fragment was determined by comparing the electrophoresed cut DNA to known standards and also observing that two highly conserved sequences were seen on the gel after electrophoresis. Through the use of ligation and transformation, DNA cloning was accomplished. Tests were run that involved electrophoresis and digestion by HindIII in order to fingerprint the DNA. The clone was identified as the desired sample through the use of transformation to identify white colonies (those with the insert) and blue colonies (those lacking the insert). The DNA molecule desired was then isolated through the use of a special resin that binds to DNA at high salt and not at low salt. The DNA is purified and the cloned sample is obtained. To verify that the cloned sample contains the correct ligating insert, digestion of the DNA was performed with HindIII and electrophoresed. The samples were found to contain the 2.0 kbp lambda fragment verifying that the sample was indeed cloned by recombinant DNA technology.
%K DNA Technology
%K Cloning
%K Genetics
%K Biotechnology
%U http://www.iiuedu.eu/press/journals/sds/SDS_2010/DSc_Article3.pdf