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Supporting File S2: Antibiotic Resistance Genes Detection in Environmental Samples - Pre- and Post-assessment multiple choice questions (DRP – ARG Assessment) DNA Structure and Replication Questions 1. What is the complement DNA strand to 5′-ATTCGGTGA-3′? A. 5′-TAAGCCACT-3′ B. 3′-CGGATTGTC-5′ C. 5′-CGGATTGTC-3′ D. 3′-TAAGCCACT-5′ E. 3-ATTCGGTGA-5 2. A DNA specimen that contains 30% guanine has ________ thymine. A. 15% B. 20% C. 30% D. 60% E. The amount of thymine cannot be determined. 3. Which of the following statements best describes DNA polymerase? A. It is an enzyme required to produce a primer needed for DNA replication. B. It is an enzyme that catalyzes the addition of nucleotides to the 5 end of a growing DNA strand. C. It is an enzyme that requires the presence of a small double-stranded nucleic acid primer to catalyze the addition of nucleotides to the 3 end of a growing DNA strand. D. It is an enzyme required to glue pieces of DNA fragments together. E. It is an enzyme that polymerizes to the DNA and keeps the replication fork open. Molecular Biology Techniques Questions 4. Only a small amount of DNA is collected from any particular soil or water sample. However, the amount of DNA collected is insufficient to perform the necessary experiments to analyze for the presence of the antibiotic resistance gene. What method could be utilized to increase the amount of DNA? A. Southern blotting B. gel electrophoresis C. polymerase chain reaction (PCR) D. colony hybridization E. DNA sequencing 5. Why is Taq polymerase required to perform a polymerase chain reaction (PCR)? 1 A. Taq polymerase is heat stable and can therefore withstand the high temperature steps required of PCR that most other enzymes cannot tolerate. B. Taq polymerase is more efficient than other polymerases. C. Taq polymerase is pressure stable and can therefore withstand the high pressure steps required of PCR that most other enzymes cannot tolerate. D. Taq polymerase makes fewer errors than other polymerases. E. All of the choices are correct. 6. Following isolation of the selected DNA fragments generated by PCR, what procedure would be used to isolate DNA fragments of different lengths? A. transformation B. transfection C. gel electrophoresis D. colony hybridization E. Western blotting 7. If a researcher used Escherichia coli DNA polymerase instead of Thermus aquaticus DNA polymerase in the PCR procedure, what would be the result? A. DNA replication would occur more slowly than normal. B. DNA replication would not occur at all. C. Many mistakes would occur. D. DNA replication would occur twice as fast as normal. E. DNA replication would stop after one cycle. 8. Which of the following would be an appropriate sequence of temperatures for PCR? A. 94°C, 55°C, 37°C B. 94°C, 65°C, 72°C C. 65°C, 72°C, 94°C D. 94°C, 37°C, 55°C E. 72°C, 65°C, 94°C 9. In gel electrophoresis, DNA molecules move toward the __________ electrode because they have an overall __________ charge. A. negative, negative B. positive, positive C. negative, positive D. positive, negative E. negative and positive, neutral 10. If all the following DNA fragments were analyzed on an electrophoresis gel, which one would migrate farthest from the negative electrode? A. 5000 base pairs 2 B. 750 base pairs C. 1000 base pairs D. 250 base pairs E. 2500 base pairs Microbiology Questions 11. Which of the following is true regarding transformed cells that are plated on growth media containing ampicillin? A. Each colony began with one antibiotic resistant cell. B. All cells in a single colony are resistant to the antibiotic ampicillin. C. Only the founder cell of a single colony is resistant to the antibiotic ampicillin. D. Cells that did not take up the plasmid will survive on the medium. E. Each colony began with one antibiotic resistant cell and all cells in the colony are resistant to the antibiotic ampicillin. 12. Which of the following statements concerning development of antibiotic resistance is FALSE? A. It is often mediated by R-plasmids. B. Resistant cells are normally in the minority in a bacterial population. C. Resistant cells grow more efficiently and quickly than susceptible cells. D. New resistance genes can be gained through transformation, transduction, or conjugation. E. Resistance can occur through mutation of existing bacterial genes. 13. Most broad-spectrum antibiotics act by __________________. A. inhibiting the synthesis of the cell wall. B. inhibiting protein synthesis. C. inhibiting nucleic acid synthesis. D. inhibiting metabolic pathways. E. disrupting the cytoplasmic membrane. 14. Drugs that affect the bacterial cell wall are active only against what type of cells? A. Gram negative B. Dormant C. Young, actively growing D. Endospore-forming E. Gram variable 15. Strategies for combating drug resistance include all but which practice? A. Responsible and appropriate drug therapy prescribed by medical personnel B. Limitation of non-medical uses of antibiotics in humans 3 C. Responsible use of drugs by patients D. Restrictions on the use of antibiotics in animal feeds E. Distribution of free prescription drugs to underdeveloped countries Data Interpretation Questions: 16. Look at the figure below and answer the following question based on your interpretation of the figure. What is the approximate size of the AmpC β-lactamase gene amplified using the blaFOX 1 primer? A. 1500bp B. 500bp C. 200bp D. Have no idea E. Impossible to determine J Clin Microbiol. 2002 June; 40(6): 2153–2162. 17. In this gel, the lanes of the gel are labeled with the name of the organism used as the source of template DNA. What is the most likely explanation for the lack of band observed when the researcher used a DNA template from K. pneumoniae, E. coli, Pseudomonas aeruginosa, S. marcescens, P. mirabilis, or E. aerogenes? Provide the most coherent explanation. A. B. C. D. Gene is not present in these organisms There was an error in the reaction set up The primers were incorrect All of the above From: J Clin Microbiol. 2002 June; 40(6): 2153–2162. Answer Key: 1D, 2B, 3C, 4A, 5C, 6C, 7E, 8E, 9B, 10D, 11D, 12C, 13B, 14C, 15E, 16E, 17C, 18A 4