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Lecture

Polymerase Chain Reaction

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Department
Biochemistry
Course
Biochemistry 2280A
Professor
Eric Ball
Semester
Fall

Description
Polymerase Chain ReactionPCR exponential amplification of any DNA from a source in which it is found as little as once involves the repeated extension of two primers that flank YFGRequired reagents for PCR 1Template DNA 2Two oligonucleotide primers which flank YFG 3dNTPs 4DNA polymerase3 Steps fig 10151 Heat to separate Strands 95 degrees C 2 Hybridization of Primers 50 degrees C going to be slightly below the Tm otherwise you would only get 50 of hybridization oligo are in range of 20 basesnot very long 3 72 degrees Cextend primers by adding DNA polymerase and dNTPSeach of the primers extends to the end of the DNA start with one molecule of template and end up with 2 Repeat this over and overfirst cycle 2 doublestranded DNA molecules Second cycleproducing 4 Thi
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