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Final-06a - ,2006 1.a.. ingthenucleotides?(4pts...

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Bio 324 Molecular Biology Final December 14, 2006 This exam is worth a total of 140 points 1. a. Nucleic acids are polymers of nucleotides. What is the name of the bond connect- ing the nucleotides? Draw the chemical structure of this bond. (4 pts.) Phosphodiester bond...sugar - o - p - o sugar b. What is one advantage of using a cloning strategy in which the foreign DNA is inser- ted into the vector using two different restriction enzymes? (4 pts.) -to efficiently amplify the sequence cheaply and accurately -inefficient to go from DNA in a test tube and have it taken up by a living cell cell have barriers c. What is the difference between a genomic and a cDNA library? If I wished to de- termine the location of introns in a given gene, what is the most reliable method to do so? (4 pts.) genomic library - contains everything, introns, exons, house keeping genes, etc cDNA - contains the coding region, so it is only exons Find the specific gene, based off of the cDNA, from there compare that region with the genomic library to find the introns d. Microarray technology is a method used to determine the expression patterns of thousands of genes at the same time. Descibe how the method works, including what is spotted on the “chip” and how the probe is generated. (4 pts.) chip is made with single stranded oligonucleotides that is 22 nucleotides long that cor- responds to sequence of known genes in a given organism. All the potential RNAs needed to be produced as single stranded or cDNA are spotted onto the chip, then isol- ate RNA from two different sources. Now the RNA has to be converted to DNA. Which can be done either by use of reverse transcriptase or amplification step using a primer to make more of the RNA. Use primers that have two different fluorescent tags. Result- ing in cDNa with two different fluorescent dyes and then mix together and hybridize. Then wash off and scan via laser 2. a. Bacterial plasmids are the most common “vector” used to clone fragments of double stranded DNA. Such plasmids often carry an antibiotic resistance gene and a
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gene encoding b -galactosidase within the multiple cloning site. Explain how each facilit- ates obtaining the desired clone. (4 pts.) Antibiotic resisteance gene allows to select only bacteria transformed by plasmid carry- ing resistence gene when plated in appropriate plates containing an antibiotic B-galactosidase which coverts x-gal to blue wil be disrupted if foreign DNA inserted into polylinker. Therefore plasmid with insert will be white in X-gal plate. b. Bacteria that lack the enzyme topoisomerase I have difficulty replicating their DNA. What is the role of topoisomerase in cells and how would that affect replication? (4 pts.) Topoisomerase relaxes the DNA by breaking bonds in DNa so that replication can oc- cur. If Topoisomerases are not present then replication cannot occur.
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