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BIS 101 Notes 2-14-11

BIS 101 Notes 2-14-11 - 3 DNA polymerase I removes RNA at...

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BIS 101 2/14/11 Lecture 11 Chapter Review Session Tue 2-22, 6-8pm in 180 Med Sci C I. DNA Structure a. DNA is the transforming agent b. The phage genetic material is DNA c. Structure of the four DNA nucleotides 1. % A = % T 2. % C = % G II. DNA Replication a. DNA is copied by semiconservative replication b. Semiconservative model predicts that half of a new strand DNA will be newly synthesized and the other is from an old strand of DNA c. Reaction is catalyzed by DNA polymerase d. DNA replication occurs at the growing fork (replisome moves alongside the growing fork) e. RNA primer synthesizes from 5’ 3’ beginning at the origin of replication 1. Before the origin of replication, strands are called the lagging strand 2. After the origin of replication, strands are called the leading strand f. Synthesizing the lagging strand 1. Primase synthesizes short RNA oligonucleotides (primer) copied from DNA 2. DNA polymerase III elongates RNA primers with new DNA
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Unformatted text preview: 3. DNA polymerase I removes RNA at 5’ end of neighboring fragment and fills gap 4. DNA ligase connects adjacent fragments g. Helicase breaks the hydrogen bonds between the two strands h. Topoisomerase controls the tension along the molecule while the strands unwind i. DNA gyrase removes extra twists III. Polymerase Chain Reaction 1. Heat to separate the strands 2. Add synthetic oligonucleotide primers and cool it down 3. Add heat tolerant DNA polymerase to catalyze 5’ 3’ DNA synthesis 4. Repeat steps 1 and 2 5. DNA synthesis (step 3) is catalyzed by the heat-tolerant DNA polymerase. 6. Repeat steps 1 through 3 IV. Dideoxy Sequencing a. 2’,3’-dideoxynucleotides cannot form a phosphodiester bond with next incoming dNTP b. It is much like a normal nucleotide, but it is missing oxygen at the 3’ and 5’ carbon, so the replication is terminated...
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