Recombinant DNA

Constructing and Screening a Recombinant DNA Library

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Session Overview

RecombDNA_sess3.jpg

This session will review how to make a recombinant genomic DNA library and how to use this library to find a specific gene. This session will outline using a library to clone a gene by complementation of a mutant phenotype.

Learning Objectives

  • To understand what a recombinant genomic DNA library is and how it is constructed.
  • To identify how to use a genomic library to identify a gene of interest.
  • To understand how to clone the gene of interest by complementation of a mutant phenoptype.
  • To understand the limitations of a genomic library.

Session Activities

Lecture Video

Watch the lecture video excerpt

Check Yourself

Question 1

Put the following steps in the appropriate order for constructing a genomic library: 1. Ligate the vector and insert DNA together using DNA ligase. 2. Isolate genomic DNA from organism. 3. Transform a population of host cells (E. coli). 4. Cut both the genomic DNA and the chosen vector with the same restriction enzyme(s). 5. Plate the transformed cells onto selective media.

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b-d-a-c-e check
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Check

Question 2

You plan to create an E. Coli genomic DNA library and clone a gene for arginine synthesis (the ARG1 gene) by complementation of a mutant phenotype. Which of the following statements is true? 1. You would isolate genomic DNA from a wild type cell. 2. You would isolate genomic DNA from an ARG1- mutant cell. 3. You would transform a population of wild type host cells. 4. You would transform a population of ARG1- mutant cells. 5. You would plate the transformed cells onto media with ampicillin. 6. You would plate the transformed cells onto media without arginine.

All of these statements are true close
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a, d, f check
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Check

Session Activities

Help Session Video

Watch the short video of Robert Dorkin explaining genomic libraries and cDNA libraries.

Practice Problems

Further Study

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Learning Resource Types

theaters Recitation Videos
assignment_turned_in Problem Sets with Solutions
grading Exams with Solutions
theaters Lecture Videos