Essential Elements of Biochemistry BCHM 3050 Dr Srikripa Chandrasekaran Lecture 2 27 15 Notes DNA Replication Part 3 I II 3 Main Steps of DNA Replication A Helicase B RNA primase DNA polymerase C Ter sequences Ter binding proteins Telomeres telomerases Polymerase Chain Reaction PCR A PCR was discovered in the 1980s by Dr Kary Mullis B 3 Main Steps of PCR 1 Denaturation 2 Annealing Heat to about 90 95 C for 1 3 minutes This separates the double stranded DNA into single strands Decrease temperature to about 40 50 C Add 2 primers template DBA dNTPs ATP CTP TTP GTP a buffer and DNA polymerase Taq DNA polymerase III cannot tolerate super high temperatures so Mullis had to find a different DNA polymerase that could tolerate the high temperatures and Taq can 3 Extension Temperature at about 70 C for 5 minutes Add the dNTPs III C 2n replication pattern Designing primers A 3 GGCC 5 primer 5 AATT gene CCGG 3 3 TTAA gene GGCC 5 5 AATT 3 primer B Primers bind to 3 end of original parent strand C Leading and lagging strands are both made new both new daughter strands 1
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