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Polymerase Chain Reaction

Polymerase Chain Reaction. DNA Structure: Two strands in helix. Adenine ( A ) binds to Thymine ( T ), with two hydrogen bonds. Guanine ( G ) binds to Cytosine ( C ), with three hydrogen bonds. DNA. Helix is double-stranded & “antiparallel” Primary strand (sense)

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Polymerase Chain Reaction

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  1. Polymerase Chain Reaction

  2. DNA Structure:Two strands in helix Adenine ( A ) binds to Thymine ( T ), with two hydrogen bonds. Guanine ( G ) binds to Cytosine ( C ), with three hydrogen bonds.

  3. DNA Helix is double-stranded & “antiparallel” • Primary strand (sense) 5’- CCGAATGGGATGC -3’ 3’- GGCTTACCCTACG -5’ • Complementary strand (antisense)

  4. PCR- Recap • Using a specific cocktail and varying temperatures/times… • TARGET & AMPLIFY desired DNA segment

  5. PCR Quiz • What are the 3 stages? • Temperatures? • What’s Happening? • In the end: How do you know if it worked?

  6. The temp cycle of PCR

  7. How Does a Gel Work? • Check a PCR sample by gel electrophoresis. +

  8. How Do you make a gel? 1% gel: Mix 0.4 grams agarose with 40ml 1X TBE buffer and heat (add 1 ul 10mg/ml ETBR stain right before pouring into tray)

  9. What’s the story, the hypothesis and was it supported?

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