Reading Guide: 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?