MCB 2007 October 10th Lecture Notes

MCB 2007 October 10th Lecture Notes - October 10th Lecture...

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October 10 th Lecture Notes: Recombinant DNA technologies continued… o Kary Mullis – a method of cloning and amplifying a sequence from vanishingly small quantities of DNA or RNA accomplished using a thermostable DNA polymerase (Taq polymerase) Polymerase Chain Reaction (PCR) Denaturation step (heating the strand, breaks the hydrogen bonds) Annealing Step (cools, allows primers to anneal to the template strand) Extension step (polymerase extends the primer using free nucleotides) components of a PCR: template DNA, forward and reverse primers, free nucleotides, Taq polymerase)
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beginning in the third cycle, amplified sequences begin to accumulate exponentially growth drops off towards the end of 30 cycles or so…chain reaction is limited by the amount of free nucleotides and stability of the polymerase (will begin to denature after a long time) can be used to look at the evolutionary relationships between organisms…sequencing following amplification
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MCB 2007 October 10th Lecture Notes - October 10th Lecture...

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