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Transcript
DNA replication, Polymorphisms, PCR and Sequencing PCR, RFLP, STR, Sequencing, SNP review 1. What is the structure of DNA? Include the parts & bonds that form a. Single nucleotide b. Backbone c. Base pairs d. Double helix e. Antiparallel f. Difference between RNA & DNA nucleotides g. Significance of the 1st, 3rd, 5th carbons in sugar (label on structure) 2. What is DNA replication? a. What are the steps b. Explain each of the following: i. DNA polymerase ii. Primase iii. Helicase iv. Ligase v. Origin of replication vi. Impact the antiparellel strands has on DNA replication vii. Nucleotides viii. Template 3. Polymorphism a. What is a polymorphism in the DNA? b. What are RFLP, STR, and SNPs? i. Include what the letters stand for, and the definition (be sure to explain the difference between each). ii. What are the limitations of each? iii. Which is the most commonly used today? Why? c. If 2 individuals had 2 different RFLPs, how would their DNA look on a gel? i. How would you prepare the DNA? d. If 2 individuals had 2 different STRs, how would their DNA look on a gel? i. How would you prepare the DNA? e. If 2 individuals had 2 different SNPs, how would their DNA look on a gel? i. How would you prepare the DNA? 4. What is PCR? a. Explain the job of each of the following: i. primer ii. taq polymerase iii. ~97 C iv. ~76 C v. ~56 C vi. chelex vii. DNA nucleotides viii. Thermocycler b. What is the purpose of PCR? c. What cell mechanisms does PCR utilize? d. Explain the steps in one cycle of PCR. e. Explain each of the reagents needed for PCR. f. How would each of the genotypes look on a gel? i. Homozygous recessive ii. Homozygous dominant iii. Heterozygous or hybrid iv. If the gene was linked to a recessive disease what would this tell you about each individuals phenotype? What would it tell you about their possibility of passing it on? g. modified from http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/R/RFLPs.html h. Using the diagram above, how many different polymorphisms are there for this gene locus? What is each person’s genotype? i. Using the diagram above, if individuals 1, 2, 6, 8 all have the same disease, would you be able to test for the disease? Why or why not? j. If individual #9 & 10 were just born what would you tell their parents? i. Would your answer change if #9 & 10 were not genetically related to this family? Why or why not? ii. Based on the genetic information above, who are possible parents of 9 & 10? Could you use this as a paternity test? Why or why not?
