Reading Guide: Pratt and Cornely, Chapter 3.1-3.3
1. Define purine, pyrimidine, nucleoside, and nucleotide.
2. From memory, draw AMP and dTTP.
3. What are other functions of nucleotides other than building blocks of DNA and RNA?
4. Draw the DNA strand dAdTdC. Label the 5’ and 3’ ends. Label the phosphodiester bond.
5. Draw the AT base pair and indicate hydrogen bonding. Do the same for the GC base pair.
6. Describe “typical” DNA (B-DNA form) in terms of its major features. How is A-DNA different?
7. What is the major structural difference between a nucleotide and a deoxynucleotide? What is the major structural difference between DNA and RNA? What is the major functional difference between DNA and RNA?
8. True or false: GC rich DNA strands are harder to separate because GC pairs form more H-bonds. Explain.
9. Draw a DNA melting curve for a helix with Tm = 78 oC. Label the y-axis and x-axis.
10. Define denature, renature, and anneal.
11. Draw a schematic for the central dogma of molecular biology, defining replication, transcription, and translation.
12. Below is the template strand for a small gene. Draw the coding strand, the mRNA produced, and the polypeptide. (Refer to Table 3.3)
3’-TCCGTAACC-5’
13. What protein is affected by the Cystic Fibrosis gene mutation? How is the protein affected?
14. About how many genes are found in E. coli? In Yeast? In humans?
15. What percentage of the human genome contains genes which encode protein products?
16. Describe two ways in which genes are identified within a genome.
Optional Lecture: Pratt and Cornely, Chapter 3.4
1. What is the goal of DNA sequencing?
2. Why is a DNA primer needed in the Sanger method? What is the role of a DNA polymerase? What is the role of a ddNTP?
3. How is pyrosequencing used to sequence a DNA strand?
4. If a DNA strand is too long for the Sanger method, how can it be sequenced?
5. What is the goal of the PCR technique?
6. Summarize the steps of the PCR reaction.
7. Define endonuclease and restriction digest.
8. What is recombinant DNA technology? What is a plasmid, and how is it different than a cloning vector?
9. How are ampR and lacZ used to select for bacteria containing a cloning vector?
10 What is site-directed mutagenesis, and how is it accomplished?
11. What are potential benefits and problems with transgenic crops?
12. What are the major problems facing greater use of human gene therapy?