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Lecture 27

BIOL 303 Lecture 27: Chapter10

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Biological Sciences
BIOL 303
Jeffrey Bell

BIOL 303 10 March 2017 “Chapter 10” Polymerase Chain Reaction (PCR) ● DNA amplification technique: ○ Uses DNA polymerase to rapidly replicate a specific DNA sequence in a test tube ● Yields more than 10 billion copies 1. DNA 2. Anneal two promers - short single strands of DNA ~20 nts a. Primers bind to each strand flanking the target b. There has to be two going inward the opposite direction 3. Extend primers w/ Taq: a. Taq is a DNA polymerase that can handle high tempature (isolated from hot springs) b. Heat temperature water loving bacteria 4. Denature DNA Products 5. 6. Extend 7. Repeat 8. Repeat again 9. 9 and again Until… ● Denature DNA with heat - 92 degrees, 30” ● Anneal primers (short DNA fragments 10-30 nucleotides long) - 50 degrees, 1’ ● Extend DNA with Taq DNA Polymerase: ○ -72 degrees, 1’ ● Repeat 30 times ● Have a million copies of the region between primers ● Produces over a million copies and separate them out “Sequencing DNA” ● Sanger sequencing: ○ Used to: ■ Sequence individual genes ■ Check the accuracy of a selected sequenced area of a genome ○ Deduces a DNA sequence by aligning pieces that differ from each other by the end base: ■ Variations label, cut, and/or immobilize the DNA pieces in different ways, g
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