Bacterial Transformation;DNA Extraction Part I & II:Total Genomic Extraction & Plasmid Extraction;Electrophoresis
By:Chris Foster
Abstract: We conducted three experiments that included a Bacterial Transformation, a two process DNA extraction, and a final procedure using gel electrophoresis. The Bacterial Transformation lab was performed to prepare the plasmid into a bacteria and to use that bacteria to amplify the plasmid in order to make large quantities of it. This is based on the natural function of a plasmid to transfer genetic information vital to the survival of the bacteria. The next process of DNA extraction involved the extraction of total genomic DNA, and extraction of bacterial plasmids. …show more content…
The gel electrophoresis portion of the lab is used to separate the DNA by size or charge. DNA Gel electrophoresis is performed for analytical purposes of the bacterial DNA used throughout this experiment.
Introduction: In a series of three lab experiments, we involved three separate linked experiments that started with transforming E.coli bacteria with a special plasmid called the pGLO (+ &-). This is a specialized plasmid that contains an abnormal promoter that is linked to GFP gene from the Aequorea victoria, a jellyfish that contains a transgenic plasmid containing DNA from two.different organisms. We want to speed up the natural process of bacterial transformation weakening the cell membranes of the bacteria through the transformation, transduction, and conjugation of the E.coli. Following this first process of the three step experiment we will being doing the first DNA extraction (Total Genomic Extraction) which involves the goal of separating the DNA from other materials so that they are readily available for further research. We will extract the DNA from our pGLO plasmid (+/-) to be used later on in our experiment …show more content…
In the Bacterial Transformation portion of the lab, we successfully weakened the bacterial cell membranes of the plasmids. In an article by Dipshikha Chakravortty, a similar bacterial transformation experiment was done using controlled micro-shockwaes to develop a unique bacterial transformation. The conditions were optimized for the maximum transformation efficiency in E.coli. This experiment was similar to the one we performed in that each experiment successfully proved that transformation can be sped up and achieved through various methods . Wanshen Yang wrote an article that uses “A multifunctional magnetic nanoparticle (MNP)-assisted bio separation method was developed to isolate plasmid DNA(pDNA) from Escherichia coli culture. Using the pH-sensitive carboxyl-modified magnetic nanoparticles, both cell capture and the subsequent removal of genomic DNA/protein complex after lysis can be achieved simply by magnetic separation.” This method is cost effective, and does not require centrifugation or precipitation steps and has the potential for automated DNA extraction. The process we used was not as efficient as this method but more cost effective in obtaining the same results. The gel electrophoresis portion of this lab was the worst of all three portions, due to