Welcome 1 2 Our goals for today Review

Welcome! • 1. 2. Our goals for today Review DNA structure and DNA amplification concepts. Amplify and analyze DNA (to investigate the use of DNA evidence as a forensics tool).

Chromosome, Gene & DNA

DNA replication

DNA replication

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Basic PCR Program Denaturation 95 degrees Annealing 65 degrees Elongation 72 degrees

What goes in a PCR reaction? Hinton Scholars AP Biology • #hinton 16

What goes in a PCR reaction? DNA Template Primers Polymerase Nucleotides Hinton Scholars AP Biology • #hinton 16

DNA analysis experimentation chain

1). DNA extraction

Lab Procedures 1. Rub the flat end of a toothpick on the inside of your cheek. 2. Twirl the end of the toothpick in 50µl X-Tract buffer. 3. Heat for 10 minutes at 95˚C. 1. DNA extraction

2). PCR

Lab Procedures 2. PCR Label the tube! Change tips every time! 3. Gently mix the reagents by pipetting up and down 3 -4 times (no bubbles), cap the tubes (firmly and tightly) 4. Place the tubes inside the PCR machine Use pipette well!

2. PCR

Lab Procedures 5. Setup the mini. PCR program (see details in the handout, remember to name and save properly!) 6. Run. (DO NOT TOUCH THE mini. PCR LID WHEN IT’S RUNNING! – HOT!) 2. PCR

3). Gel electrophoresis

DNA gel electrophoresis Use electrical field to pull DNA to migrate from one end to the other end of the gel Power source ? ? 1. DNA is negatively charged 2. DNA travel through the pores (in gels) --- “slowly” 3. The speed that is inversely related to their lengths (molecular weight).

Lab Procedures 2. Gel electrophoresis 1. Place the completely submerged into the electrophoresis buffer (no bubbles in the wells). Release the DNA slowly into the wells! 5. Visualize the DNA

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Positive Negative control Hinton Scholars AP Biology • #hinton 16