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You want to select two restriction enzymes that would be beneficial in the identification of pMICO. You have decided that Notl will be one of the enzymes you will use. Which restriction enzyme buffer will you use in the digest of pMICO? Select one: a. Restriction Buffer B b. Restriction Buffer D Correct c. Restriction Buffer C d. Restriction Buffer E e. Restriction Buffer A b. Restriction Buffer D You want to select two restriction enzymes that would be beneficial in the identification of pMICO. You have decided that Notl will be one of the enzymes you will use and want to find another enzyme that would be compatible in a double digest. Given the list of available restriction enzymes, which other enzyme cuts pMICO once and uses the same Restriction buffer? Select one: a. Sca1 b. Nru1 c. Hpa1 d. Sac11 e. Sal1 Correct f. Sac1 g. BamH1 h. Spe1 i. Bcl1 j. EcoR1 e. Sal1 Restriction enzymes have an optimal incubation temperature which is defined in the enzyme manufacturer's instructions. What is the optimal incubation temperature to digest pMICO with the restriction enzymes selected in questions 8 and 9? Select one: a. 37ºF This is incorrect. Please refer to the Promega website following the link provided in the Practical Manual. b. 42ºC c. 54ºC d. 37ºC d. 37ºC What size fragments would you expect if you digested pMICO in: 1: a single digest with restriction enzyme 1 2: a single digest with restriction enzyme 2 3: a double digest with restriction enzymes 1 and 2 Select one: a. 1: 6652 bp, 2: 6652 bp, 3: 2933 & 3719 bp correct b. 1: 1620 bp, 2: 4553 bp, 3: 2933 & 3719 bp c. 1: 6652 bp, 2: 6652 bp, 3: 479, 1620 & 4553 bp d. 1: 1620 bp, 2:4553 bp, 3: 479, 1620 & 4553 bp a. 1: 6652 bp, 2: 6652 bp, 3: 2933 & 3719 bp

You want to set up a restriction enzyme digest of your isolated plasmid, pMICO. You determine that the concentration of your plasmid DNA is 50 nanograms/microlitre. How many microlitres of your purified plasmid DNA would you add to a 20 microlitre digest, in order to add 250 nanograms DNA to the digest? You do not need to include units in your answer. Answer: Feedback Correct The correct answer is: 5 The correct answer is: 5 You want to set up a single restriction enzyme digest of your isolated plasmid, pMICO. Given that you have included 250 nanograms of your plasmid DNA (refer to question 12) and the correct volumes of 10x Restriction Buffer + 10x BSA, how much distilled water would you add to a 20 microlitre digest? Select one: a. 9 μl b. 11 μl c. 20 μl d. 2 μl e. 10 μl e. 10 μl You want to set up a double restriction enzyme digest of your isolated plasmid, pMICO. Given that you have included 250 nanograms of your plasmid DNA (refer to question 12), and the correct volumes of 10x Restriction Buffer + 10x BSA, how much distilled water would you add to a 20 microlitre digest? Select one: a. 11 μl b. 2 μl c. 10 μl d. 9 μl Correct. e. 20 μl d. 9 μl You want to set up a control (no restriction enzyme added) for the digest of your isolated plasmid, pMICO. Given that you have included 250 nanograms of your plasmid DNA (refer to question 12) and the correct volumes of 10x Restriction Buffer + 10x BSA, how much distilled water would you add to a 20 microlitre digest? Select one: a. 10 μl b. 20 μl c. 9 μl d. 11 μl Correct e. 2 μl d. 11 μl How many grams of agarose would you add to 40 millilitres of 1 x TAE to prepare a 0.8% agarose gel? Select one: a. 3.2 grams b. 8 grams c. 0.8 grams

d. 0.32 grams Correct e. 10 grams d. 0.32 grams...