BIOL 222 Lecture 8: PCR and DNA Sequencing

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Pcr = polymerase chain reaction: developed in 1985 by kary mullis, method you can amplify (copy) a speci c region of dna. Ingredients for pcr: template dna (of what you want to copy), polymerase, dntps (all four nucleotides), and primers. Dntp = dttp, datp, dctp, and dgtp. Two kinds of primers are used forward primer and a reverse primer: bind to opposite sides of the template strands, functions of these ingredients. The two primers provide a free 3" end for polymerase to bind to: these primers also have a speci city on where they bind, which allows us to control what the polymerase will copy. Steps of pcr: 1. heat the template strands to 95 c to denature and separate the two strands by breaking the hydrogen bonds. In humans this role is done by helicase: 2. primer annealing. 1min/thousand bp ~72 : 4. repeat (denature, anneal, extend)

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