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.
Watch the lecture video excerpt
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.
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.
Help Session Video
Watch the short video of Robert Dorkin explaining genomic libraries and cDNA libraries.