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PCR & Sequencing

PCR & Sequencing. polymerase chain reaction ( PCR). produce many copies of a specific target segment of DNA. thermocycler. dNTPs (free nucleotides) Taq polymerase buffer 2 DNA primers. Figure 13.25. 2. 1. 3. Technique. 3 . 5 . PCR

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PCR & Sequencing

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  1. PCR & Sequencing

  2. polymerase chain reaction(PCR) • produce many copies of a specific target segment of DNA

  3. thermocycler • dNTPs(free nucleotides) • Taq polymerase • buffer • 2 DNA primers

  4. Figure 13.25 2 1 3 Technique 3 5 PCR http://www.dnalc.org/resources/animations/pcr.html PCR song: ~2min http://www.cnpg.com/video/flatfiles/539/ Target sequence Genomic DNA 3 5 3 5 Denaturation 5 3 Annealing Cycle 1 yields 2 molecules Primers Extension New nucleotides Cycle 2 yields 4 molecules Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence

  5. DNA Sequencing • Relatively short DNA fragments • Can be sequenced by the Sanger dideoxychain-termination method http://www.dnalc.org/resources/animations/cycseq.html

  6. HumanGenomeProject 1990-2003 Craig Venter’sshotgun-sequncing-1000bp per day in 1980s-1000bp per second in 2000-100 billionbp per day in 2010!

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