prelim2.key - BioBM332 Second prelim 3/13/08 Name: _KEY_ 1....

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BioBM332 Second prelim 3/13/08 Name: __________KEY______________________ 1. _______ 2. _______ 3. _______ 4. _______ 5. _______ 6. _______ Total _______ 1
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1. (18 points) In the Sanger sequencing reaction, using dideoxynucleotides was the key to this reaction. How is the dideoxynucleotide differed from deoxynucleotides in a) chemical structure? ___3'-OH in deoxynucleotides and 3'-H in dideoxynucleotide in the 5C sugar 3 pts b) in the polymerization reaction ? dideoxynucleotides are terminators because the 3'-H cannot form a phosphoester bond with the 5'-P of the incoming nucleotides 3 pts The following DNA fragment was sequenced by the Sanger method. The asterick indicates a fluorescent label. *5’ 3’OH 3’ ATTGTACATGGC---------5’ A sample of the DNA was reacted with DNA polymerase and each of the nucleotide mixtures (in appropriate buffer) listed below. a. All 4 dNTPs and ddATP in relatively small amounts b. All 4 dNTPs and ddCTP in relatively small amounts c. dATP, dTTP and dCTP only d. All 4 dNTPs and all 4 ddNTPs in vast excess e. All 4 dNTPs and all 4 ddNTPs in relatively small amounts The resulting DNA samples were loaded onto wells in lanes a-e respectively and separated by electrophoresis on an agarose gel. Indicate in the gel below the fluorescent band pattern in each of the lanes. Write the sequence deduced from the reaction on the side of the gel.
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This note was uploaded on 03/30/2008 for the course BIOBM 3320 taught by Professor Tye,bk during the Summer '07 term at Cornell University (Engineering School).

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prelim2.key - BioBM332 Second prelim 3/13/08 Name: _KEY_ 1....

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