BIO250L+V2+Lab+7 straighterline assignment

lOMoARcPSD| BIO250L+V2+Lab+7 straighterline assignment Human Biology Lab ( ) lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L Student Name: Access Code (located on the underside of the lid of your lab kit): AC-XC9QOE Pre-Lab Questions 1. Which DNA nitrogenous bases pair with each other? Which bases are purines, and which are pyrimidines? Guanine pairs with cytosine. Adenine pairs with Thymine. Purine: adenine and thymine Pyrimidines: guanine and cytosine 2. How is DNA information used to make proteins? What are the steps of this process? Enzymes read information in DNA molecules, then it's transcribed to mRNA and the information from the mRNA is translated through tRNA and created with amino acids to build the 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. PCR is used to amplify small DNA samples. Recombinant DNA technology is used to alter DNA. 4. What occurs during each of the three steps involved in the PCR cycle? How has the use of PCR changed biotechnology? Denaturation is separating DNA. Annealing is when primers attach to DNA. Extensions are newly created DNA. 5. How could you take a protein with a known sequence of amino acids and use it to create an artificial gene? lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L Use recombinant DNA to alter the sequence of the amino acid to create the 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 DNA phosphate charge b. Detergent (in the DNA extraction solution): release DNA from the membrane c. Ethanol: promotes ionic bonding and turns DNA into beads 2. What else might be in the ethanol/aqueous interface? How could you eliminate this? DNA & RNA which can be removed through centerfiguing. 3. What is the texture and consistency of the DNA? Thin, fragine, long 4. Is the DNA soluble in the aqueous solution or in the alcohol? DNA is soluble in the aqueous solution. Insert a photo of your DNA. Include your name and access code handwritten in the background of lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L your photo. 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 2810 Post-Lab Questions 1. What is the expected size of the plasmid plus the cut foreign DNA? 3,530 2. What type of ends do the enzymes BamHI and EcoRI produce? How does this type of end facilitate cloning? The cloning process is facilitated by the enzymes producing bonds with sticky ends. lOMoARcPSD| Lab 7 Microbial Genetics & Genetic Engineering BIO250L 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 because the enzymes use ATP to create bonds. 4. How would you clone a gene into a plasmid if there were no common restriction sites between the two DNA sequences? The gene could be cloned through the process of electroporation if there were no common restriction sites between the DNA sequences.