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Review Chapter 3 and 4 Matching For the following items, match each word with the most appropriate definition below. a. The proportion of solute to solvent in a solution b. The concentration of a solution expressed as the number of moles of solute per liter of solvent c. A solution that resists changes in ph d. What gets dissolved in a solution e. The number of molecules that equals the molecular weight of a substance, in grams f. The addition of solvent to make a mixture less concentrated g. The solution that dissolves added molecules h. Containing water 1. aqueous 2. buffer 3. concentration 4. dilution 5. molarity 6. a mole 7. solute 8. solvent 9. 50 µl 10. 3.5 g 11. 3 µl 12. 0.07 mg 13. 4 ml 14. 200 µl 15. 1.5 L 16. 32 g For the following items, select which tool below would most accurately measure the following amounts. a. micropipet b. pipet c. graduated cylinder d. tabletop balance e. analytical balance

In the following items, determine which equation (a-c) would be most useful in calculating the amount of solute needed to make each solution. a. Concentration Volume = Mass b. Decimal Value Volume = Mass c. Volume Molarity Molecular Weight = Mass 17. 1 L of 3 M NaOH 18. 250 µl of 0.08 g/ml hemoglobin solution 19. 10 ml of 3% saline solution 20. 600 µl of 0.5 mm protein solution 21. Base Pair 22. Enhancer 23. Exon For the following items, match each word with the most appropriate definition (a-j) below. a. A region on prokaryotic DNA where one or more genes and their controlling elements are located b. Process of correcting DNA codes that cause genetic diseases and disorders c. Cells that have taken up foreign DNA and start expressing it d. Source of nutrients, in liquid or solid form, for growing cell cultures e. A region of DNA that can increase expression of a gene f. Using electricity to separate molecules on a gel slab g. Two nitrogenous bases connected with hydrogen bonds h. A DNA molecule that is used as a host to carry foreign pieces of DNA into a cell i. The expressed part of genes j. Non-coding intervening sequence within genes 24. Gel electrophoresis 25. Gene therapy 26. Intron 27. Media 28. Operon 29. Transformed 30. Vector For the following items, place the following steps of genetic engineering in the proper order. a. First b. Second c. Third d. Fourth 31. Transfer gene of interest into a host organism

32. Harvest desired product from cell culture 33. Isolate gene of interest 34. Identify a molecule that can be improved/enhanced For the following items, which gel electrophoresis system best matches the following descriptions or applications? a. Agarose b. Polyacrylamide c. Either 35. Used for separating proteins 36. Used in vertical gel boxes 37. Used in horizontal gel boxes 38. Used to separate DNA samples 39. Separates larger molecules such as restriction digest fragments 40. Separates smaller molecules such as polypeptides Multiple Choice Identify the choice that best completes the statement or answers the question. 41. Which of the following can be measured using a micropipet? a. A gram of sodium chloride b. A liter of oxygen gas c. 10 µl of DNA solution d. 2.5 L of protein solution 42. 240 µl is equivalent to a. 2.4 ml. b. 0.24 ml. c. 0.024 ml. d. 0.0024 ml. 43. When using a micropipet, the second stop on the plunger is used to a. eject the micropipet tip into the waste container. b. evacuate all the air in the micropipet and draw in the volume indicated on the display. c. evacuate all the air in the micropipet when dispensing a reagent. d. prevent cross-contamination if you forget to change pipet tips. 44. As a solution becomes more concentrated, a. the proportion of solute to solvent increases. b. the proportion of solvent to solute increases. c. the normality of the solution shifts. d. deionization begins to occur. 45. Which of the following is an expression of concentration units?

a. µg/µl b. mg/ml c. µm d. All of the above 46. How much solute is needed to make 550 ml of 4 g/l gelatin solution? a. 220 mg b. 0.72 g c. 2.2 g d. 8 g 47. How much protein do you need to add to a 250 µl reaction that requires a concentration of 8 mg/ml? a. 0.2 ng b. 2.0 ng c. 0.2 mg d. 2.0 mg 48. 3.3 liters is equivalent to a. 3300 ml b. 330 ml c. 33 ml d. 0.03 ml 49. How much 95% ethanol do you need to make a 600 ml solution of 70% ethanol? a. 1.1 L b. 81.4 ml c. 110 ml d. 442 ml 50. As a solution becomes more dilute a. the proportion of solute to solvent increases. b. the proportion of solvent to solute increases. c. the normality of the solution shifts. d. re-ionization begins to occur. 51. A buffer is a solution that a. decreases the ph and alters the physical structure and function of molecules in that solution. b. resists a change in ph and preserves the structure and function of molecules in that solution. c. increases the ph and alters the function of molecules. d. alters the ph of a reaction but maintains the structure and function of molecules. 52. A p100 micropipet is used to measure volumes a. under 100 ml. b. over 100 ml. c. between 10 and 100 µl. d. over 100 µl. 53. A p1000 micropipet is used to measure volumes a. between 100 and 1000 µl. b. between 1-1000 µl. c. over 1000 µl.

d. between 10-1000 ml. 54. A p10 micropipet is used to measure volumes a. between 0.1 and 10 µl. b. between 1 and 10 µl. c. between 10 and 100 µl. d. over 10 µl. 55. What is the equivalent of 0.75 ml? a. 7.5 µl b. 75 µl c. 750 µl d. 7500 µl 56. How much 50X TAE buffer would you need to make a 2 liter solution of 1X TAE buffer? a. 40 ml b. 100 ml c. 200 ml d. 1 L 57. 300 ml of a 1/40 dilution of 20X TBE buffer stock solution is needed, what volume of solute is needed? a. 2.7 ml b. 7.5 ml c. 40 ml d. 150 ml 58. When measuring liquids using a graduated cylinder or pipet, a. the top of the meniscus should be at the graduation. b. the bottom of the meniscus should be at the graduation. c. the meniscus should be anywhere near the graduation. d. there is no meniscus when using these measurement tools. 59. What would be the most accurate way to dispense 64 sets of 15 µl? a. Use a micropipet in each hand. b. Use a pipet and add one drop to each tube. c. Use a multi-channel micropipet. d. Use a regular micropipet. 60. If a volume you wish to measure is within the range of two different micropipets, which would give the most accurate measurement? a. The lower range micropipet is more accurate. b. The higher range micropipet is more accurate. c. They are equally accurate. d. There are no micropipets whose ranges overlap. 61. In a biotechnology lab, tap water is best used for a. making buffers. b. washing glassware. c. the final rinse when washing glassware. d. All of the above 62. Which solution is more concentrated? a. 5 mg/ml

b. 5 µg/µl c. 5 g/l d. They are equivalent. 63. What is the lowest setting on a p10 micropipet? a. 0.1 µl b. 0.5 µl c. 1.0 µl d. 10 µl 64. Calculate the mass of 17 moles of HCl. a. 17 amu b. 170 amu c. 306 g d. 620 g 65. Calculate the mass of 0.3 moles of NaOH. a. 8.4 g b. 12 g c. 3.6 amu d. 28 amu 66. The order of amino acids on a polypeptide chain is encoded by a. DNA. b. mrna. c. rrna. d. regulatory proteins. 67. Nucleotides are connected together in a DNA molecule by a. hydrogen bonds. b. antiparallel bonds. c. peptide bonds. d. phosphodiester bonds. 68. How is gene expression controlled in prokaryotic cells? a. Operons b. Ribosomes c. Enhancers d. All of the above 69. All of the following may be used to induce changes at a particular locus on a DNA molecule except: a. Exposure to chemicals b. Exposure to radiation c. Exposure to low temperatures d. Viral infection 70. How many hydrogen bonds hold base pairs together? a. Two hydrogen bonds for each pair b. Three hydrogen bonds for each pair c. Two hydrogen bonds between A and T, and three for G and C d. Three hydrogen bonds for A and T, and two for G and C. 71. How often will cells divide under ideal growing conditions?

a. Every 20 minutes b. Every 45 minutes c. Every 2 hours d. Every 12 hours 72. How do DNA molecules vary from one species to another? a. The type of nitrogenous bases they contain b. Number of genes and non-coding regions c. The way nitrogenous bases pair d. The directionality of the strands 73. All of the following regulate gene expression in eukaryotes except: a. Enhancers b. Transcription factors c. Histone-DNA complexes d. Operators 74. What is a possible outcome of site-specific mutagenesis? a. Additions or deletions to DNA b. Improved protein function c. Cell death d. All of the above 75. All of the following are reasons some people need recombinant protein therapeutics except: a. They need stronger nails and hair b. Their body doesn t synthesize that protein at the right time c. The protein their body synthesizes is defective d. Their body doesn t synthesize enough of that protein 76. Prokaryotic DNA is contained in a. the cytoplasm. b. the nucleus. c. membrane-bound organelles. d. multiple chromosomes. 77. Which of the following is an example of sterile technique? a. Using distilled water b. Treating a lab bench with disinfectant c. Using dust-free glassware from the glass storage shelves d. Rinsing Petri dishes in warm water 78. A typical E. coli genome contains about genes. a. 2 b. 5-10 c. 2000 d. 4000 79. In eukaryotic cells, after an mrna has been transcribed, a. the operon turns off. b. it immediately gets translated into a polypeptide. c. exons are removed. d. introns are removed.

80. All of the following are reasons to use TRIS during an electrophoresis except a. To stabilize the ph b. To conduct electricity c. To maintain the shape of molecules d. To stain molecules 81. R plasmids are useful for both bacteria and scientists because they contain a. ribosomal RNA genes. b. one or more antibiotic resistance genes. c. recombinant DNA products. d. All of the above 82. Which stain causes DNA to glow orange when exposed to UV light? a. Agarose b. Polyacrylamide c. Methylene blue d. Ethidium bromide 83. A circular plasmid was cut with a restriction enzyme and three bands were visualized after electrophoresis. How many cuts were made in the plasmid? a. 1 b. 2 c. 3 d. 4 84. A linear piece of DNA was cut with a restriction enzyme and four bands were visualized after electrophoresis. How many cuts were made? a. 1 b. 2 c. 3 d. 4 85. Eukaryotic gene expression can be increased by a. operators. b. introns. c. enhancer molecules. d. All of the above 86. Which agarose concentration would work best for separating very large molecules? a. 3% b. 2% c. 0.8% d. All of the choices would work well 87. When a regulatory molecule binds to an operator, a. gene expression is turned on. b. gene expression is turned off. c. RNA polymerase may bind to the promoter region. d. enhancer molecules fall off. 88. Most of the DNA in humans is a. coding sequence. b. spacer DNA within and between genes.

c. for regulatory processing. d. for transcription factor gene regulation. 89. What is agar? a. Solid media on which bacteria grow b. Liquid media for bacterial growth c. Region on a bacterial operon d. Matrix used for vertical gel electrophoresis 90. How is the packaging of DNA different in eukaryotes and prokaryotes? a. Eukaryotes have more chromosomes and each is linear b. Eukaryotes have more chromosomes and each is circular c. Eukaryotes have some carbohydrate molecules attached to their chromosomes. d. Eukaryotes have less chromosomes than prokaryotes. Short Answer 91. Write step-wise instructions to train a new lab technician to prepare a solution of 2 L of 50 mm. Complete the calculations and include instructions on how to use the electronic balance and how to prepare the solution. 92. You want to teach a friend how to use a micropipet. How would you instruct her so that she could correctly dispense 45 µl of plasmid DNA from one tube into another? Include instructions on how to set the volume on the micropipet, draw in and dispense the liquid, and eject the tip into the waste container. You have p20, p200, and p1000 micropipets available. 93. A lab technician diluted 25 µl of a 50X sample with enough solvent to reach a final volume of 3 ml. What is the new concentration of the sample? Show your work. 94. Indicate the pipet needed and draw the dial setting for the following volumes: a. 1.6 µl b. 45 µl c. 170 µl d. 950 µl 95. You need to dilute 50X buffer into 1 Liter of 3X buffer. In your lab, you have p10, p100, and p1000 micropipets, 5mL, 10 ml, and 25 ml pipets, 50 ml, 100 ml, 500 ml, and 1L graduated cylinders. How much stock buffer do you need and which measurement tools will you need to use? Show your work and justify your selection of measurement tools considering accuracy and efficiency. 96. List five similarities in DNA from one organism to another. 97. Describe the mechanism of an operon. 98. Describe how gene therapy is used for patients with cystic fibrosis. 99. Identify how each of the following organisms is used for the genetic engineering of tissue plasminogen activator. 100. Explain how to prepare an agarose gel electrophoresis system.

Review Chapter 3 and 4 Answer Section MATCHING 1. ANS: H PTS: 1 REF: 75 2. ANS: C PTS: 1 REF: 86 3. ANS: A PTS: 1 REF: 76 4. ANS: F PTS: 1 REF: 85 5. ANS: B PTS: 1 REF: 77 6. ANS: E PTS: 1 REF: 82 7. ANS: D PTS: 1 REF: 75 8. ANS: G PTS: 1 REF: 76 9. ANS: A PTS: 1 REF: 69 10. ANS: D PTS: 1 REF: 75 11. ANS: A PTS: 1 REF: 69 12. ANS: E PTS: 1 REF: 75 13. ANS: C PTS: 1 REF: 68 14. ANS: A PTS: 1 REF: 69 15. ANS: C PTS: 1 REF: 68 16. ANS: E PTS: 1 REF: 75 17. ANS: C PTS: 1 REF: 84 18. ANS: A PTS: 1 REF: 78 19. ANS: B PTS: 1 REF: 80 20. ANS: C PTS: 1 REF: 84 21. ANS: G PTS: 1 REF: 101 22. ANS: E PTS: 1 REF: 108 23. ANS: I PTS: 1 REF: 108 24. ANS: F PTS: 1 REF: 116 25. ANS: B PTS: 1 REF: 111 26. ANS: J PTS: 1 REF: 108 27. ANS: D PTS: 1 REF: 103 28. ANS: A PTS: 1 REF: 104 29. ANS: C PTS: 1 REF: 104 30. ANS: H PTS: 1 REF: 104 31. ANS: C PTS: 1 REF: 112-113 32. ANS: D PTS: 1 REF: 112-113 33. ANS: B PTS: 1 REF: 112-113 34. ANS: A PTS: 1 REF: 112-113 35. ANS: B PTS: 1 REF: 117 36. ANS: B PTS: 1 REF: 117

37. ANS: A PTS: 1 REF: 117 38. ANS: C PTS: 1 REF: 117 39. ANS: A PTS: 1 REF: 117 40. ANS: B PTS: 1 REF: 117 MULTIPLE CHOICE 41. ANS: C PTS: 1 REF: 85-87 42. ANS: B PTS: 1 REF: 67 43. ANS: C PTS: 1 REF: 72 44. ANS: A PTS: 1 REF: 76 45. ANS: D PTS: 1 REF: 77 46. ANS: C PTS: 1 REF: 78 47. ANS: D PTS: 1 REF: 78 48. ANS: A PTS: 1 REF: 67 49. ANS: D PTS: 1 REF: 75 50. ANS: B PTS: 1 REF: 76 51. ANS: B PTS: 1 REF: 86 52. ANS: C PTS: 1 REF: 68 72 53. ANS: A PTS: 1 REF: 73 54. ANS: B PTS: 1 REF: 73 55. ANS: C PTS: 1 REF: 68 56. ANS: A PTS: 1 REF: 87 57. ANS: B PTS: 1 REF: 87 58. ANS: B PTS: 1 REF: 68 59. ANS: C PTS: 1 REF: 73-74 60. ANS: A PTS: 1 REF: 72 61. ANS: B PTS: 1 REF: 76 62. ANS: D PTS: 1 REF: 67 63. ANS: B PTS: 1 REF: 73 64. ANS: D PTS: 1 REF: 83 periodic table 65. ANS: B PTS: 1 REF: 83 periodic table 66. ANS: B PTS: 1 REF: 99-100 67. ANS: D PTS: 1 REF: 107 68. ANS: A PTS: 1 REF: 104 69. ANS: C PTS: 1 REF: 114 70. ANS: C PTS: 1 REF: 101 71. ANS: A PTS: 1 REF: 106 72. ANS: B PTS: 1 REF: 102 73. ANS: D PTS: 1 REF: 108 74. ANS: D PTS: 1 REF: 114 75. ANS: A PTS: 1 REF: 113 76. ANS: A PTS: 1 REF: 103 77. ANS: B PTS: 1 REF: 106 78. ANS: C PTS: 1 REF: 103 79. ANS: D PTS: 1 REF: 108-109

80. ANS: D PTS: 1 REF: 118 81. ANS: B PTS: 1 REF: 103 82. ANS: D PTS: 1 REF: 118 83. ANS: C PTS: 1 REF: 119 84. ANS: C PTS: 1 REF: 119 85. ANS: C PTS: 1 REF: 108 86. ANS: C PTS: 1 REF: 118 87. ANS: B PTS: 1 REF: 105 88. ANS: B PTS: 1 REF: 108 89. ANS: A PTS: 1 REF: 105 90. ANS: A PTS: 1 REF: 103 SHORT ANSWER 91. ANS: Formula weight for = 24.31 g/mol + 2(35.45 g/mol) = 95.21 g/mol 800mL 50mMol/L 1 Mol/1000mM 95.21 g/mol 1L/1000mL= 3.81g To measure 3.81g using an electronic balance, turn power on, add weigh paper or weigh boat, press tare, wait for balance to zero, measure out 3.81 g of and add to a 1L graduated cylinder. Add deionized water until the bottom of the meniscus reaches the 800 ml mark. PTS: 1 REF: 83 92. ANS: Pick the p200 micropipet and put on a new tip, then dial 045 on the micropipet. Press the plunger down to the first stop, put the tip in the liquid, and draw in 45 µl of plasmid DNA by slowly releasing the plunger. Transfer to a new tube and press the plunger past the first stop, all the way to the second stop. This will expel all the solution from the tip. Remove the micropipet from the second tube, while sill holding the plunger down, and then release. Press the button near the plunger to eject the tip into a waste container. PTS: 1 REF: 72 93. ANS: = 50X = 25 µl = unknown = 3 ml Set up the = equation and solve for : 50X buffer 25 µl = C 2 3000 µl = 50X = 0.42X

PTS: 1 REF: 86 94. ANS: A: 1.6 µl with p10 micropipet would read 016 B: 45 µl with p100 or p200 micropipet would read 045 C: 170 µl with p200 micropipet would read 170 or on p1000 micropipet would read 017 D: 950 µl with p1000 micropipet would read 095 PTS: 1 REF: 72-74 95. ANS: = 50X = unknown = 3X = 1 L = 1000 ml Set up the = equation and solve for : 50X buffer = 3X buffer 1000 ml = 3X 1000 ml 50X =60 ml Measure 60 ml of 50X buffer into a 100 ml graduated cylinder (smallest vessel that will hold entire volume). Pour into a 1 liter graduated cylinder (smallest vessel that will hold entire solution) and bring to volume with deionized water until the bottom of the meniscus reaches the 1L mark. PTS: 1 REF: 87 96. ANS: CATEGORY 1: Contain same bases (A, C, G, T) in which A and T pair and G and C pair together. CATEGORY 2: Double helix structure held together by hydrogen bonds CATEGORY 3: Strands are antiparallel CATEGORY 4: 10 bases/turn of the helix, each is 0.34 nm apart CATEGORY 5: Replicate in a semi-conservative fashion PTS: 1 REF: 101-102 97. ANS: An operon is a unit composed of a promoter, one or more structural genes, and a terminator. An RNA polymerase binds to the promoter site to activate the gene. The enzyme works its way down towards the structural gene and then synthesizes the mrna that will then get translated into a peptide chain. Transcription can be halted if a regulatory molecule binds to the operator region, just prior to the structural gene, blocking RNA polymerase from reaching the structural gene it intended to transcribe. PTS: 1 REF: 104-105 98. ANS: A functional CFTR (cystic fibrosis transmembrane conductance regulator) gene was inserted into a modified adenovirus and delivered to the cells that line the nose.

PTS: 1 REF: 115 99. ANS: Human: Source of the t-pa gene that needs to be engineered Bacteria: Serves as vector for cell transformation CHO cells: This organism mass produces the t-pa protein PTS: 1 REF: 113 100. ANS: STEP 1: Prepare agarose and pour into plastic mold with a plastic comb. Allow it to cool so the gel solidifies. STEP 2: Place gel in the gel chamber, with the wells at the end with the negative electrode. STEP 3: Pour buffer into the gel chamber until the gel is submersed. STEP 4: Load samples into the wells of the gel. STEP 5: Plug in the electrodes and set the power supply to deliver current through the gel. STEP 6: After the molecules have migrated through the gel, stain the gel and visualize. PTS: 1 REF: 116-117

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