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Lecture7MCB160Lf11 - Mallery 2006 U Miami Thermophilus...

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Mallery 2006 U. Miami
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Thermophilus aquaticus is a source of Thermo- stable DNA Polymerase: “Taq” Polymerase Thermophilus aquaticus was first isolated from hot springs in Yellowstone National Park. The yellow patches are photosynthetic bacteria and the white areas correspond to warmer water (70°C - 80°C) that contains stringy masses of T. aquaticus (Images from Woods Hole Oceanographic Institution www.divediscover.whoi.edu)
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http://www.dnalc.org/ddnalc/resources/pcr.htm See PCR animation: Link is on course website Or use the URL below
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PCR yield and e ff٠ ciency Expected yield: 300 μ g Actual yield: usually ~1 μ g, i.e. much less than expected PCR amplification is not 100% efficient i.e. it is not 2X per cycle, but smaller. Usually from 1.75 to 1.95 per cycle, depends upon DNA sequence and size. At 90% efficiency, i.e. 1.9 per cycle 30 cycles gives (1.9) 30 = 0.2 billion fold i.e. 0.2 X theoretical maximum of 1 billion In this case, the expected yield =60 μ g (still far higher than actual yield)
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