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MOLECULAR BIOLOGY EXAM II February 19, 2003 ______________________________ Good luck! Remember, a drawing or figure is often more valuable than words. Tables are great. As always, I will "sell" you part of the answer if you need it. Make all answers fit in the spaces given. I Fill in (3 pts per space.) 1,2) 3) 4) 5,6) 7) 8) 9,10) Small non-coding RNA molecules found in the nucleus are called ___________________ and _____________________. Design a 12 base (single stranded) probe for Northern Blot, using the following amino acid sequence: leu-met-gly-phe-ser-glu-asp met-trp ____________________. ____________________ enzyme is used to make a cDNA library from the mRNA. The stringency of hybridization can be increased or decreased by altering __________________________ and/or ____________________. A screening tool in which many DNA sequences are placed on a chip for hybridization analysis. ______________________ ___________ is the term used to define an area of sequence which codes for protein, and is not interrupted by stop codons. Protein __________________ are often defined by ______________ in the gene. II 5 points each. Choose 4 to describe/draw in several sentences. linkage analysis shotgun sequencing pcr SNP microsatelites chromosome walking III 1) Essay questions 10 points Describe the three main types of RNA. Draw two of them. 2) 15 points Describe physical and genetic mapping, discussing the following; What is the basis for each type? What types of techniques are used in each type of mapping? (list several) What type of data is obtained? Why are both techniques necessary? (how do they complement each other?) 3) 10 points Describe the function of each of these enzymes, and indicate when and why you might use them in lab: DNA polymerase ligase nuclease 4) 10 points research study You have identified a new protein found in liver, and have determined its sequence. The protein is an enzyme that functions in repairing cellular damage by toxins. You have developed a simple assay to test the presence and activity of this enzyme. You know that the possibilities for using this enzyme in medicine are enormous. You need to know- What is its gene sequence? Where is it made? Do other organisms make this protein? Are the sequences from other organisms similar? Is it always made or only at certain times? How is the gene regulated? You have three people working for you, all are pretty handy in the lab. Outline a strategy for each to begin tackling one of these questions, (or another critical issue you think is important to market this protein). Be sure the three areas will provide information that is complementary, and provides important info. Outline in flow chart form how each person will achieve their goal, including brief info about techniques used, and the type of data they will obtain. You can use the back of this sheet. THINK before you write Strategize. 5) 5 points Make up and answer a very good question. If in doubt- ask me! Samples: Compare and contrast the use of RFLP and PCR analysis in forensic analysis; RNA modification; DNA purification