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) 5’- CCGAATGGGATGC -3’ 3’- GGCTTACCCTACG -5’ Complementary strand (antisense) PCR- Recap • Using a specific cocktail and varying temperatures/times… – TARGET & AMPLIFY desired DNA segment PCR Quiz • • • • What are the 3 stages? Temperatures? What’s Happening? In the end: How do you know if it worked? The temp cycle of PCR 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?