Isolation, Restriction Digestion, And Electrophoresis Of...

Isolation, restriction digestion, and gel electrophoresis of plasmid DNA Prathyusha Gudapati, BIOL 304, spring 2015. Abstract The purpose of the experiment was to isolate plasmid DNA, followed by restriction digestion using restriction endonucleases and then visualizing the digested fragments after subjecting to gel electrophoresis. Plasmid DNA (pSP72 DNA) was isolated from Escherichia coli KAM32 (E.coli) cultures using the QIA prep miniprep kit and then subjected to restriction digestion by EcoRI and HindIII. The restriction digested DNA was then loaded into the wells of 0.7% agarose gel and subjected to electrophoresis. It can be concluded from our results that our plasmid DNA isolation was successful and the restriction digestion results were partially in agreement with our hypothesis. Introduction Plasmids are small double stranded circular non chromosomal DNA molecules containing their own origin of replication. Hence, they are capable of replication independent of the chromosomal DNA in bacteria. Plasmids present in one or more copies per cell, can carry extra chromosomal DNA from one cell to another cell and serve as tools to clone and manipulate genes. Plasmids used exclusively for this purpose are known as vectors. The genes of interest can be inserted into these vector plasmids creating a recombinant plasmid. Recombinant plasmids can play a significant role in gene therapy, DNA vaccination, and drug delivery [Rapley, 2000]. We hypothesized that plasmid DNAShow MoreRelatedCreating A Genomic Library Of The Bacteria Aliivibrio Fischeri1768 Words   |  8 PagesIntroduction Within a genome, there is a vast sequence of DNA that may be studied. The resulting goal of this study is to create a genomic library of the bacteria Aliivibrio Fischeri. We will be achieving this purpose by making Escherichia Coli luminescence through the use of the lux operon. In the process of understanding the genomic library of A. Fischeri bacteria, we will be creating a restriction map of the restriction sites in the plasmids containing a lux. In this study we will be working withRead MoreManipulation And Analysis Of Dna Using Standard Molecular Biology Essay717 Words   |  3 Pagesanalysis of DNA using standard Molecular Biology Techniques. During the course of the next three practical classes you shall be performing a number of techniques in order to isolate and manipulate DNA from bacteria. The practicals are spread over three sessions, the techniques that you will perform are indicated below: Practical 1 Isolation of plasmid DNA from three cultures of E.coli using a method known as the alkaline lysis method. 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