BLD 430 Quiz 10 1.

Sequencing reactions are often performed in the same fashion as a standard PCR reaction with the incorporation or inclusion of ddNTP's into the reaction mixture. A) True B) False 2. ddNTP's are nucleotides that lack the 3'OH group necessary to allow elongation or extension of the new strand of DNA being synthesized by the polymerase enzyme. A) True B) False 3. Sequencing reactions in the clinical setting often use the amplicon or amplified fragment generated in a previous PCR reaction as a template. A) True B) False 4. Sanger, chain termination, sequencing reactions can use the same primers to initiate extension by the polymerase enzyme that were used to generate the template. A) True B) False 5. When a sequencing reaction is set up, only one primer can be used. A) True B) False 6. It is not uncommon to see heterozygote positions, two bases in the same place, on an electropherogram because we are often sequencing a heterozygous individual. A) True B) False 7. Pyrosequencing exploits the phosphorous molecules that are released in the process of adding one base to another during chain elongation by the polymerase enzyme. A) True B) False 8. Stronger signals or higher peaks are seen closer to the primer in a sequencing reaction than out near the end of a long (>500bp) sequencing run/reaction. A) True B) False 9. Single base resolution is better at the 3' end of the sequencing reaction rather than the 5' end. A) True B) False

10. Discovery of a what is suspected to be a new allele would require that the template be regenerated and re-sequenced 6-10 times to ensure accuracy as well as be sequenced in both directions (use both primers in separate reactions). A) True B) False