5.1 Manipulating DNA

Some notes adapted from Mrs. Savage Please complete accompanying question sheet and activity.

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5.1 Manipulating DNA

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Some notes adapted from Mrs. Savage Please complete accompanying question sheet and activity.
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  1. 5.1 Manipulating DNAEntrance ticket

    Slide 1 - 5.1 Manipulating DNAEntrance ticket

    • In crime shows such as CSI, the forensic techs are often taking tiny samples of evidence left at the scene and matching the DNA on them to suspects. How does this actually work?
  2. Manipulating DNA

    Slide 2 - Manipulating DNA

    • Standard 5.1
  3. Tools of DNA technology: Enzymes

    Slide 3 - Tools of DNA technology: Enzymes

    • DNA polymerase- synthesizes DNA
    • Primers- initiate DNA transcription
    • Ligase- glues ends of DNA strands together
    • Restriction endonuclease- cut DNA at specific points
  4. Restriction Enzymes

    Slide 4 - Restriction Enzymes

    • Restriction enzyme – enzymes that cut strands of DNA in particular places
    • Every restriction enzyme is programmed to cut whenever it sees a particular base pair sequence
    • Where the restriction enzyme will cut varies based on the DNA sequence
  5. Restriction Enzymes

    Slide 5 - Restriction Enzymes

    • Can create sticky ends or blunt ends
  6. Restriction Fragment Length Polymorphism (RFLP) Analysis

    Slide 6 - Restriction Fragment Length Polymorphism (RFLP) Analysis

    • Uses restriction enzymes to cut DNA at certain places and chop the DNA into pieces
    • Analyzes the length of the pieces to see how similar or different two samples of DNA are.
    • Uses gel electrophoresis to make a restriction map
    • Different individuals will have different restriction maps
  7. Gel electrophoresis

    Slide 7 - Gel electrophoresis

    • Separates pieces of DNA based on how many base pairs long they are
    • Longer pieces of DNA move through the gel slower
  8. Gel Electrophoresis

    Slide 8 - Gel Electrophoresis

    • Prepare gel
    • DNA sample cut with restriction enzyme
    • Pipette DNA into wells
    • Turn on the current
    • Stain with DNA-binding dye
    • Rinse dye
    • See results
  9. Gel Electrophoresis

    Slide 12 - Gel Electrophoresis

    • What is the purpose of gel electrophoresis?
  10. Gel Electrophoresis

    Slide 13 - Gel Electrophoresis

    • What is the purpose of gel electrophoresis?
    • To compare DNA from different sources to see how they are similar and different
    • To determine if someone has a genetic disease
    • Match DNA from crime scene to suspect (should be 100% match)
    • Test for paternity (should be 50% match)
    • Why don’t we test for maternity often?
  11. PCR:Polymerase chain reaction

    Slide 14 - PCR:Polymerase chain reaction

    • PCR can replicate specific segments of DNA many times very efficiently.
    • Why it is useful to make many copies of a small amount of DNA? In what situations would it be helpful?
  12. PCR Process

    Slide 15 - PCR Process

    • DNA is heated to separate the strands.
    • Primers bind to ends of target sequence.
    • DNA polymerase binds and copies DNA.
    • Repeat 1-3 many times.
    • Result: Many copies of a target sequence.
    • PCR video
  13. More tools: Probes

    Slide 17 - More tools: Probes

    • Complimentary pieces of DNA
    • with a stain or radioactive label
    • attached
    • Bond to specific DNA sequences
    • Helps find specific genes
  14. Tools of DNA technology: Plasmids

    Slide 19 - Tools of DNA technology: Plasmids

    • Small, circular section of DNA
    • Replicates in bacteria
    • Can carry genes
    • Can be copied from one bacterium to another