BABS1201 Lecture Notes - Lecture 17: Radiography, Pyrimidine Dimer, Reverse-Transcriptase Inhibitor
8 Jun 2018
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BABS1201 Notes – LEC 17, 18
LEC 17: PCR
• Original concept for PCR:
o Denature DNA strands just below 100℃
o Primers bind (anneal) to DNA between 70-37℃
o DNA polymerase synthesises new strand at 37℃
o Repeated
• Problem: DNA polymerase is inactivated at 90℃ which is required to denature DNA
strands
• Modified concept for PCR:
o Denature DNA strands just below 100℃
o Primers bind (anneal) to DNA between 70-37℃
o DNA polymerase from the thermophile Thermus aquaticus (Taq polymerase)
is stable at 98℃ and works at 70℃, therefore Taq DNA polymerase can
synthesise new DNA at 70℃
• Simple sequence repeats (SSRs) can accumulate mutations unhindered as they
mostly occur in non-coding regions and vary from person to person
• To amplify a single specific SSR by designing primers that bind immediately adjacent
to the SSR
• Individuals all have different SSRs therefore electrophoresis of a forensic sample
compared to suspects can determine which matches the DNA at the crime
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