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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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- Winter '08