Practice Final Examkey

Practice Final Examkey - LS3 Final Exam INSTR: L. Johnson...

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LS3 Final Exam INSTR: L. Johnson Thursday, Dec. 12, 2002 NAME:_________________________________ (last) (first) I.D. #__________________________________ INSTRUCTIONS 1. This exam is 10 pages long. PAGE VALUE SCORE Before starting make sure you have all the pages. 2 19 2. Be sure to write clearly. Only 3 12 legible answers will be graded. 4 22 5 30 6 17 7 17 8 18 9 21 10 18 11 26 200 Lab 20 Total 220
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2 NAME:_________________________________ ID# ________________________ (last) (first) Part A. New Material (100 pts) (8 pts) 1. Restriction enzymes recognize palindromic sequences and cut both strands of DNA at the same sequence. Consider the four enzyme recognition sites shown below and the types of fragments they generate. Restriction Enzyme Recognition Site BspE I 5’ T^CCGGA 3’ Eag I 5’ C^GGCCG 3’ Apa I 5’ GGGCC^C 3’ NgoM I 5’ G^CCGGC 3’ a) Which enzymes produce a 4 base sticky ends? All of the enzymes shown above. (4 pts) b) If you wanted to clone a fragment which was digested with one enzyme into a vector digested with another enzyme, which two enzymes above could be used (i.e. have compatible ends for cloning)? BspE I and NgoM I (4 pts) No partial credit for either answer shown above. (11 pts) 2. a) Hybridization is used in many molecular biology techniques. Why is this such a powerful technique? It allows one to detect a unique DNA molecule amongst a mixture of molecules. It is the basis of Southern analysis, northern analysis, colony and plaque hybridization and many other techniques. b) Describe in general what an antibiotic is and how it is used during cloning procedures. An antibiotic is a compound that can specifically inhibit an enzyme required for cell function. It is used in cloning to select for the presence of vectors that carry genes the render the cell resistant to the antibiotic, in other words it is used as a selectable marker. c) Describe in general what an antibody is and how it is used during cloning procedures. An antibody is a protein made in eukaryotic cells that binds tightly to an antigen and signals an immune response. It can be used to screen expression libraries for the synthesis of a particular protein, allowing the appropriate clone to be recognized.
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3 AME:_________________________________ ID# ________________________ (last) (first) (12 pts) 3. You have succeeded in isolating a novel human virus and proceed to create a restriction map using two enzymes, Sal I and Bam HI. After digestion and electrophoresis you obtain the results shown below. However, when you try to determine the restriction map you find there are two possible maps consistent with the data you obtained. To distinguish which map is correct, you repeat the digestions and electrophoresis, but this time you transfer the NA to nitrocellulose and perform a Southern analysis, using the 4 kb restriction fragment from the Bam HI + Sal I digest as a probe. Using both sets of data, determine the correct restriction map. Agarose Gel
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Practice Final Examkey - LS3 Final Exam INSTR: L. Johnson...

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