BIOA01H3 Midterm: Module 2 Review

Order the following steps of the DNA replication process fromfirst to last. (From 1 to 8)

DNA ligase joins Okazaki fragmments together

Elongation of leading strands occurs continuosly and elongationof lagging strands occurs via generation of Okazaki fragments. Bothare synthesized in 5' to 3' direction.

DNA polymerase I removes the RNA primers and replaces them withDNA nucleotides

DNA polymerase III starts adding DNA nucleotides to the 3' endof the existing RNA chain.

Helicase untwists the double helix at the replication fork

Single-stranded binding proteins bind the unpaired DNA strands,keeping them from re-pairing.

Topoisomerase helps releive the strain caused by DNA untwisting.It breaks, swivels and rejoins DNA strands.

Primase starts a complementary RNA chain using parental DNAstrand as a template

1) What is the difference between the leading strand and the lagging strand in DNA replication? Place the following steps of DNA replication in right chronological order.

1. Primase adds an ribonucleotide primer
2. Initiator proteins bind the origin of replication and helicase breaks hydrogen bonds between complementary bases of double stranded template DNA
3. DNA polymerase III extends by complementary base pairing deoxyribonucleotides to the template strands
4. Ligase phosphodiester bonds the nucleotides to connect okazaki fragments nad connect fragments at the origin and where the "crotches or forks" collide
5. DNA polymerase I removes RNA primer and replaces with Deoxyribonucleotides

2) Given the template DNA and primer sequences below, what bases will be added to the primer as DNA replication proceeds? (The bases should appear in the order that they will be added)

5'-GTCGTCTCCCTTTTTAAAAACCCCCCC-3'
5'-UUUUUAAAAAGGG-3'


3) Given the DNA sequence below, what could be a primer sequence used in its replication?
5'-CGTACGGCCCCAAAATTGCTACTCTGCTG-3'

PCR Amplification, sequencing and Genetic identification of Prokaryotes

1. Compare DNA replication in a typical cell to the synthetic DNA replication that happens during the Polymerase Chain Reaction (PCR). Fill in the missing information that describes how each process is accomplished (i.e. enzyme name or other method) in each. Two answers are given as example:

2. Heating proteins to 94°C would destroy the function of most human or bacterial enzymes, including DNA Polymerase. How is this avoided during the DNA strand synthesis step in PCR?

3. What are the two major factors that influence the rate at which DNA amplicons move through an electrophoresis gel?

4. What are the three most significant features of the 16S rRNA gene that make it an excellent choice for DNA homology analysis and identification in bacteria?