Lab 7 for Microbiology Lab from Straighterline

lOMoARcPSD| Lab 7 for Microbiology Lab from Straighterline Fundmtls Of Microbiology W Lab lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L” Student Name: Access Code (located on the underside of the lid of your lab kit): “Pre-Lab Questions” 1. Which DNA nitrogenous bases pair with each other? Which bases are purines, and which are pyrimidines? Adenine pairs with Thymine and Cytosine pairs with Guanine. Adenine and Guanine are both purines and Thymine and Cytosine are pyrimidines. 2. How is DNA information used to make proteins? What are the steps of this process? DNA information is used to make proteins through transcription and translation. The first step is when the enzymes from the DNA molecules are transcribed into mRNA. The mRNA goes to the ribosome creating tRNA and is translated into amino acids that block proteins. 3. Give an example of a scenario in which you would perform PCR vs a scenario in which you would use recombinant DNA technology. A scenario when I would need to perform PCR is when determining the source of a disease or illness and collecting DNA from blood samples. When you’d have to use recombinant DNA technology would be when needing to take human growth hormones. 4. What occurs during each of the three steps involved in the PCR cycle? How has the use of PCR changed biotechnology? The three steps involved in the PCR cycle are denaturing, annealing, and elongation. Denaturing is when the DNA is heated up and separates into 2 single strands. Annealing is the next step, temperatures cool down and the DNA primers attach themselves to the DNA template. In elongation, the temperature goes back up high and the new strand of lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L” DNA is made. The use of PCR has changed biotechnology because it can now be applied medically and is used to identify pathogens. 5. How could you take a protein with a known sequence of amino acids and use it to create an artificial gene? To create an artificial gene, you can use recombinant DNA technology like bioinformatic tools. Having a protein that has a known sequence of amino acids can create an artificial gene. “EXPERIMENT 1: DNA Extraction POST-LAB QUESTIONS 1. What is the purpose of the following reagents in the experiment? a. Salt (in the DNA extraction solution): Neutralizes the charge of DNA’s phosphate, enables precipitation, and allows the DNA to attach b. Detergent (in the DNA extraction solution): Breaks the membrane and releases DNA, RNA, and proteins. c. Ethanol: Promotes iconic bonds and precipitation, overall causing the DNA to separate from the solution. 2. What else might be in the ethanol/aqueous interface? How could you eliminate this? RNA and Soluble salt might be in the ethanol/aqueous interface. This can be eliminated by using the centrifuge. 3. What is the texture and consistency of the DNA? It is slimy, stringy, and fragile. lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L” 4. Is the DNA soluble in the aqueous solution or in the alcohol? Soluble in the aqueous solution. Insert a photo of your DNA. Include your name and access code handwritten in the background of your photo. lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L” “EXPERIMENT 2: Cloning a DNA Fragment to a Bacterially-Derived Plasmid Vector DATA TABLES Table 1: Fragment Lengths DNA Type Longest Length (in base pairs) Foreign 720 Plasmid 2508 POST-LAB QUESTIONS 1. What is the expected size of the plasmid plus the cut foreign DNA? The expected size of the plasmid plus the cut foreign DNA is 3,228 base pairs. 2. What type of ends do the enzymes BamHI and EcoRI produce? How does this type of end facilitate cloning? The enzymes BamHI and EcoRI form double bonds and they produce sticky ends that help facilitate the process of cloning. The plasmid DNA can be linked by DNA ligase in the correct orientation in the host genomic DNA. 3. What enzyme is necessary to permanently link the digested foreign and plasmid DNA together to form the recombinant DNA molecule? How does this enzyme work? Ligase is an enzyme that is necessary to permanently link the digested foreign and plasmid DNA together that then forms a recombinant DNA molecule. Ligase is necessary because it uses ATP to form covalent bonds and links the phosphate sticky end to the DNA strand sticky end. 4. How would you clone a gene into a plasmid if there were no common restriction sites between the two DNA sequences? lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L” If there were no common restriction sites between the two DNA sequences, you would clone a gene into the plasmid by electroporation.