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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

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
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 2 transformation
Step 2: Transformation

Introduction of the vector+insert into a host

  • Common example = electroporation
step 2 transformation1
Step 2: Transformation

Selecting cells with a vector

Selectable

marker

step 2 transformation2
Step 2: Transformation

Selecting cells with a vector

step 2 transformation3
Step 2: Transformation

Distinguishing insert-containing vectors

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

slide14

cDNA

Library

slide15

cDNA

Library