Monday october 13 2008 chapter 9 part ii of iii
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Monday, October 13, 2008 Chapter 9, Part II (of III) PowerPoint PPT Presentation

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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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


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


Step 1: Ligation of DNA into vector


Different types of vectors


Step 2: Transformation

Introduction of the vector+insert into a host

  • Common example = electroporation


Step 2: Transformation

Selecting cells with a vector

Selectable

marker


Step 2: Transformation

Selecting cells with a vector


Step 2: Transformation

Distinguishing insert-containing vectors

(if the process worked perfectly, this wouldn’t be necessary)


Cloning review


Purifying the cloned DNA


Separating insert from vector


Subcloning (from vector to vector)


Genomic Library


cDNA

Library


cDNA

Library


Genomicvs. cDNA


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