Gene Cloning And Bacterial Transformation

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Introduction: In this lab, gene cloning and bacterial transformation was conducted. Bacteria are single celled microorganisms. They are simple in structure and have no nucleus with linear chromosomes. Bacteria are essential to life, and reproduction of other bacteria which helps play a role in decomposition of organic matter and the cycling of elements such as nitrogen and carbon, which are elements necessary to live. Due to this, plants and animals can use nitrogen to create nucleic acids along with amino acids, which are the essential building blocks of DNA. They also help break down food in the digestive system, which allows us to obtain the nutrients we need, and bacteria helps develop our immune system to fight other illnesses that…show more content…
The bacterium being observed was E. coli, which codes for green fluorescent protein. E. coli is a bacterium that normally lives in the intestines of humans and animals alike, and can cause infection. One hopes to observe a glow from a green fluorescent protein (GFP) which is typically made by a jellyfish. This will be made possible by the additional presence of arabinose, which acts as the inducing agent. The inducing agent influences the transcription of the GFP and activates the transcriptional promoters. A promoter allows a foreign gene to fall into place much easier resulting in gene expression. It is expected that the combination of the plasmid, with the Lysogeny Broth (LB) and the arabinose (ARB) will transform the bacteria, and allow for growth of the bacteria in the presence of the antibiotic ampicillin (AMP) along with an observable glow under ultraviolet (UV) light. Hypothesis: • The Agar plate with the LB/AMP/ARB in the +pGLO will be the only plate to display growth and a glow when compared to the other agar plates. Materials: • Water bath set at 42 degrees Celsius • Pipette tips • Micropipetter (P20) • 50 mM CaCl2 solution • pGLO plasmid • Sterile loop • Competent E. coli HB101 • 37-degree incubator • Sterile LB agar plates (LB, LB/amp, and LB/amp/ara) • Sterile transfer pipette Methods: 1. After labeling each of the tubes, 250 mL of CaCl2 was added to all the tubes and
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