Plasmid

Transformation of pGlO plasmid to an E.coli bacteria cell We pipetted 250 ul of CaCl2 transformation solution to into two separate tubes. Next, we used a sterile loop to transfer two to four larger bacteria colonies from an E.coli plate into each of the two tubes. The bacteria was observed under UV light to make initial observations. We pipetted 10 ul of pGLO plasmid (0.08 ug/ul) into one of the tubes. Both tubes were incubated on ice for ten minutes. After ten minutes of incubation, the tubes…

AIM: To create competent E.coli cells by using the chemical method (calcium chloride treatment) To transform the competent bacterial cells using plasmid DNA and evaluate the transformation efficiency. PRINCIPLE: Competent cells are pre-prepared bacterial cells that possess altered cell walls through which foreign DNA can enter the cell easily. Most cells cannot take up DNA efficiently unless they have been treated with chemicals or exposed to electrical treatments in order to make them…

in transformations since the effects will be seen much faster than with other hosts. When the bacterium receives the genes from the plasmids, it provides better traits for adaptation and survival, but it is only successful if all substances needed are in the mixture as well (Masterman 2010). My hypothesis is that using the heat shock method described above, the plasmid pGLO will be used as a vector to transfer green fluorescent protein, arabinose C, and ampicillin resistance from the pGLO to the…

method called heat shock. Prokaryotic cells that have been artificially transformed by the foreign DNA can then be calculated for efficiency of transformation. pGLO plasmid was an excellent choice of foreign DNA, due to its ability to alter the phenotype of the prokaryotic cell. Green fluorescent protein (GPF gene) found in the plasmid must be exposed to arabinose, a sugar, in order to activate the GPF gene. If transformation occurred within the prokaryotic cell, the cell will fluoresce…

Polymerase chain reaction (PCR) Generalities PCR is a method developed in 1985 by Kary Mullis and to obtain, by coupling a heat-resistant DNA polymerase and without cloning, amplification of a fragment of DNA known. Initially, DNA polymerase was isolated from a bacterium thermophilne (resistant to significant increases in temperature), as Thermus aquaticus (Taq polymerase). Currently, we use recombinant enzymes, whose; elaboration is easier and greater efficiency. The general principle of PCR is…

Change for a Longer Life Span Not a Better Span of Life. Everyone has different beliefs and thoughts about genetically modified organisms. Many believe that it is okay as long as the individuals ends up better than they were while others believe that an infant child should not be altered for any reason. What if a change in their genome can cause them to be immune to certain diseases, or even help cognitively with dyslexia? Would people still be opposed to something that can help an individual…

known as, Recombinant DNA. Recombinant DNA, otherwise known as rDNA, is an artificial DNA strand, created by combining gene sequences with one another. In order to create a rDNA you must first isolate the DNA donor, as well as the vector. Bacterial plasmids are commonly used, so for this explanation it will be used as the vector. The next step is to cut the DNA. This is possible with the help of restriction enzymes, which act as scissors cutting only at certain DNA targets sequences (restriction…

in Escherichia coli (E. coli) by isolating and purifying a plasmid from E. coli, determining its orientation with respect to the plasmid backbone by restriction mapping, and transforming it into an alternate E. coli strain. Methods: Each individual received either E. coli A or B strain of pKN800 plasmid DNA. The plasmid was purified and isolated, then cut with PstI restriction enzyme to create a sample of PstI-cut and uncut pKN800 plasmid DNA and to distinguish orientation A from B. These…

the new trait has been passed on. Bacteria naturally contain one or more small circular pieces of DNA called plasmids. Plasmid DNA usually contains genes for one or more traits that may be beneficial to bacterial survival. In nature, bacteria can transfer plasmids back and forth, allowing them to share…

World Book Student defines genetic engineering as “techniques that alter the genes (hereditary material) or combination of genes in an organism… Beginning in the 1970’s, scientists developed ways to reintroduce individual genes into cells or into plants, animals, or other organisms” (Rubenstein, 2015). Genetic engineering is a controversial subject not only in science but also in popular culture. Some research, such as stem cell research, has led to serious questions concerning ethics. Other…