lab-7-microbial-genetics-genetic-engineering

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Studocu is not sponsored or endorsed by any college or university LAB 7 Microbial Genetics & Genetic Engineering Human Resources (Mohammad Ali Jinnah University) Studocu is not sponsored or endorsed by any college or university LAB 7 Microbial Genetics & Genetic Engineering Human Resources (Mohammad Ali Jinnah University) Downloaded by Brenda Ledbetter (bledbetter0627@gmail.com) lOMoARcPSD|28593260
Lab 7 Microbial Genetics & Genetic Engineering BIO250L ” Student Name: Hannah Ejalonibu Access Code (located on the underside of the lid of your lab kit): AC-VGS01NE “ Pre-Lab Questons ” 1. Which DNA nitrogenous bases pair with each other? Which bases are purines, and which are pyrimidines? The nitrogenous bases pair are :Cytosine and Thymine both of them are the pyrimidines,Adenine and guanine both of them are the purines. 1. How is DNA informaton used to make proteins? What are the steps of this process? First of all enzymes read the information in a DNA molecule and transcribe it into an intermediary molecule which is mRNA (messenger RNA). The information contained in the messenger RNA molecule is translated for amino acids. 2. Give an example of a scenario in which you would perform PCR vs a scenario in which you would use recombinant DNA technology. For PCR, it is when I have an unknown DNA isolated from a specimen in small concentration, and I want to amplify it so it can be identified through gel electrophoresis.To identify the function of a gene for DNA technology it would be. 3. What occurs during each of the three steps involved in the PCR cycle? How has the use of PCR changed biotechnology? For any DNA synthesis reaction PCR is required 3 steps first one is Downloaded by Brenda Ledbetter (bledbetter0627@gmail.com) lOMoARcPSD|28593260
Lab 7 Microbial Genetics & Genetic Engineering BIO250L ” denaturation of the template into single strands second one is annealing of primers to each original strand for new strand synthesis and third one is extension of the new DNA strands from the primers. 4. How could you take a protein with a known sequence of amino acids and use it to create an artfcial gene? I can make its most probable nucleotide sequence based on the frequency of tRNA that is present in the organism in which i need to express the proteins. By using PCR, i will then artificially make the sequence and amplify it. “ EXPERIMENT 1: DNA Extracton Post-Lab Questions 1. What is the purpose of the following reagents in the experiment? a. Salt (in the DNA extracton soluton): Salts helps to make debris like proteins, RNA clump together and lipids. b. Detergent (in the DNA extracton soluton): The purpose of detergent is to pulls the nucleus releasing the DNA from the cell and the membrane surrounding the cells. c. Ethanol: Ethanol purpose is to cause precipitation and promotes ionic bonds. Downloaded by Brenda Ledbetter (bledbetter0627@gmail.com) lOMoARcPSD|28593260
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Lab 7 Microbial Genetics & Genetic Engineering BIO250L ” 2. What else might be in the ethanol/aqueous interface? How could you eliminate this? RNA and soluble salt .By using a RNase enzymes ,centrifuge , or crushing it in a bag and draining it through a cheese cloth it could be eliminated . 3. What is the texture and consistency of the DNA? Sticky slimey and Fragile 4. Is the DNA soluble in the aqueous soluton or in the alcohol? aqueous solution. Insert a photo of your DNA. Include your name and access code handwriten in the background of your photo. “ EXPERIMENT 2: Cloning a DNA Fragment to a Bacterially-Derived Plasmid Vector Downloaded by Brenda Ledbetter (bledbetter0627@gmail.com) lOMoARcPSD|28593260
Lab 7 Microbial Genetics & Genetic Engineering BIO250L ” Data Tables Table 1: Fragment Lengths DNA Type Longest Length (in base pairs) Foreign 726 Plasmid 2823 Post-Lab Questions 1. What is the expected size of the plasmid plus the cut foreign DNA? 3,549. 2. 3. What type of ends do the enzymes BamHI and EcoRI produce? How does this type of end facilitate cloning? Sticky ends that help facilitate the cloning process. Plasmid DNA can be linked by DNA ligase in the correct orientation in the host genomic DNA 4. 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 because the ligase use ATP to form covalent bond and link the phosphate sticky end to the DNA strand sticky end 5. How would you clone a gene into a plasmid if there were no common restricton sites between the two DNA sequences? I would clone a gene by using electroporation. Downloaded by Brenda Ledbetter (bledbetter0627@gmail.com) lOMoARcPSD|28593260

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