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Monday, October 13, 2008 Chapter 9, Part II (of III)

Monday, October 13, 2008 Chapter 9, Part II (of III). Assignment : Chapter 9 problems 1-4, 6, 9, 16, 20, 25, 26, 32 Chapter 7 problems due this week and QUIZ!. Cloning. Molecular Cloning. Step 1: Ligation of DNA into vectors Sticky ends Different types of vectors Step 2: Transformation

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Monday, October 13, 2008 Chapter 9, Part II (of III)

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  1. Monday, October 13, 2008Chapter 9, Part II (of III) Assignment: Chapter 9 problems 1-4, 6, 9, 16, 20, 25, 26, 32 Chapter 7 problems due this week and QUIZ! Cloning

  2. Molecular Cloning • Step 1: Ligation of DNA into vectors • Sticky ends • Different types of vectors • Step 2: Transformation • Putting recombinant DNA into host cell • Selecting the cells with inserted DNA • Distinguishing insert-containing vectors • Purifying the cloned DNA • Genomic and cDNA libraries

  3. Step 1: Ligation of DNA into vector

  4. Different types of vectors

  5. Step 2: Transformation Introduction of the vector+insert into a host • Common example = electroporation

  6. Step 2: Transformation Selecting cells with a vector Selectable marker

  7. Step 2: Transformation Selecting cells with a vector

  8. Step 2: Transformation Distinguishing insert-containing vectors (if the process worked perfectly, this wouldn’t be necessary)

  9. Cloning review

  10. Purifying the cloned DNA

  11. Separating insert from vector

  12. Subcloning (from vector to vector)

  13. Genomic Library

  14. cDNA Library

  15. cDNA Library

  16. Genomicvs. cDNA

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