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LAB 6 - LAB6STUDYGUIDE KaryMullis: Bacteria/ temperatures...

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LAB 6  – STUDY GUIDE Kary Mullis:  awarded Nobel Prize in Chemistry for development of PCR Critical component of PCR – getting DNA Polymerase from  Thermophilic  organism Bacteria/Eukarya that grow at high temperatures – DNA polymerase works at high  temperatures Process of PCR: 1. Thermostable DNA polymerase, nucleotides, and DNA are heated in a solutions  containing high concentrations of primers 2. Primers are complementary to specific DNA sequences 3. As mixture cools, primers hybridize to DNA  4. Thermostable DNA polymerase uses nucleotides to copy DNA strand 5’ to 3’ 5. First cycle gets you 2 new copies from two original strand templates 6. Mixture is heated so strands of DNA are separated but polymerase isn’t affected 7. Solution is cooled again to allow primers to hybridize to the 4 strands – repeat 5 minute  cycles to get lots of amplification Cycles: 1. 95 ° C – DNA strand denaturation 2. 58 ° C – Primer annealing 3. 72 ° C – DNA elongation Components: DNA Template 2 Primers (5’ and 3’) DNA Polymerase dNTP’s Buffer
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