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DNA Technology

DNA Technology. Copying DNA: PCR. Polymerase Chain Reaction Gene Amplification A method of making many copies of a piece of DNA. PCR: Copying DNA. DNA, nucleotides, buffers, “taq” polymerase, and two primers are placed in a small test tube taq polymerase can work at very high temps.

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DNA Technology

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  1. DNA Technology

  2. Copying DNA: PCR • Polymerase Chain Reaction • Gene Amplification • A method of making many copies of a piece of DNA

  3. PCR: Copying DNA • DNA, nucleotides, buffers, “taq” polymerase, and two primers are placed in a small test tube • taq polymerase can work at very high temps

  4. PCR: Step 1 • The DNA is heated (80oC), the two strands separate • Primers match with complementary bases • Taq pol. begins adding new nucleotides (5’->3’)

  5. PCR: Step 2 • The tube is cooled the DNA strands together • Cycle repeated to make several copies

  6. PCR Large amounts of DNA can be made from a small starting sample

  7. Cloning Just a gene vs. whole organims

  8. Bacterial plasmids in gene cloning

  9. Cloning a gene • Requires: • Plasmid (cloning vector), restriction enzymes, and gene of interest • DNA ligase attaches the sticky ends of DNA fragments together • Chimeric DNA made!

  10. Cloning an organism • A body cell from one organism and an egg cell from another are fused • The resulting cell divides like a normal embryo

  11. Cloning “Dolly”

  12. Cutting DNA • Restriction enzymescut DNA at specific sequences • Useful to divide DNA into manageable fragments

  13. Electrophoresis • DNA can be separated based on size and charge • The phosphate groups are negatively charged • DNA is placed in a gel and electricity is run through

  14. Electrophoresis • Negative DNA moves toward the positive end • Smaller fragments move farther and faster

  15. Electrophoresis

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