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Gene Cloning Part 1: The Mechanics of Recombinant DNA
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Gene Cloning Part 1: The Mechanics of Recombinant DNA
Authors
Don Lee, Department of Agronomy and Horticulture, University of Nebraska-Lincoln
Patricia Hain, Department of Agronomy and Horticulture, University of Nebraska-Lincoln
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Gene Cloning Part 1: The Mechanics of Recombinant DNA
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Objectives for Gene Cloning Part 1
The Gene Cloning Process
Why clone genes?
DNA is cloned from DNA
Genes are Connected to Other Genes
Gene Cloning Requires Knowledge of the Gene’s Unique Characteristics.
Plasmids do the “Tricking”
Cloning with LacZ and X-gal plasmids
Step One: Make Sticky Ends
Step Two: Make Recombinant Plasmids
Step Three: Transformation of E. coli
Step Four: Selecting Bacteria with Recombinant Plasmids
Step Five: Making Libraries of Genes
Genomic Libraries
cDNA library
Gene Libraries with Big Pieces
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