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ANSWERS
The number of bases per turn of DNA is___________
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~10.5
What does the Meselson and Stahl experiment confirm?
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DNA replication is semi-conservative
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Heavy and light strands with nitrogen isotopes prove
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True or False|\ |\
a. Most biological DNA is positively supercoiled.
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b. Triplex DNA is pyrimidine rich
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c. Chargaff's rule: A=G & T=C
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d. A single zinc finger can recognize 10bp of DNA
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A. False, negatively = Left-handed. Some is right handed =
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positive
B. True
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C. False A=T C=G
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D. False, 3 bp, use multiple zinc fingers in a row for more
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sequence specificity/bp |\
,DNA is not a uniform structure, many localized variants
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can substitute protein domains, alpha helix/beta hairpin, in zinc
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fingers to make hybrids with different binding specificities
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(designated by alpha helix) for major grooves of DNA |\ |\ |\ |\ |\ |\ |\ |\
x-ray crystallography allows us to view and design custom
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designer DNA-binding proteins = alter gene fxn in intact cells
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In A & Z DNA:
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a. Which is right handed helix?
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b. Which one has 12bp per helical turn?
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c. Which is the dehydrated form?
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a. A
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b. Z
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c. A
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primary DNA is B |\ |\ |\
Gel electrophoresis
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molecular separation technique |\ |\
,can separate nucleic acids or proteins
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make AGAROSE gel with slots
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hot liquid poured with "teeth"
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put DNA in slot, run electric current at neutral pH
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DNA = negatively charged (phosphates) = migrates towards
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positive end |\
key is friction
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DNA stained with fluorescent dye, looked at under UV
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unknown DNA electrophoresed parallel with known to determine
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size
distance migrated = can plot on graph with log of molecular
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weights or number of bp |\ |\ |\ |\
ex: 20mm = 910bp
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PFGE = Pulsed Field Gel Electrophoresis
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for longer sequences
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, as large as some chromosomes in yeast
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What is PAGE? |\ |\
Polyacrylamide Gel Electrophoresis |\ |\
used for PROTEINS |\ |\
have to denature the protein = treat with detergent, SDS, sodium
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dodecyl sulfate
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SDS-PAGE
SDS advantages:|\
1) coats polypeptides with (-) charge
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2) masks natural charge so results are purely based on molecular
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weight
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What is 2-D gel electrophoresis?
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LESS POWER: use two different pH values to compare, pH affects
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net charge so different rates of mobility
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