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BIO SCI 99 Study Guide - Midterm Guide: Reverse Genetics, Oligonucleotide, Cloning

Biological Sciences
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Lecture 13 Molecular Cloning, cDNA Library and Functional Complementation
(Greyed-out text is meant only to be helpful and is optional)
1. Molecular Cloning
What is molecular cloning?
Basically refers to the process of creating a DNA sequence of interest and inserting it
into some kind of vector for the purpose of introducing this DNA into a cell for
production of the gene product (if the DNA of interest is a gene) or to genetically
manipulate the cell for studying cellular pathways and gene function.
What are the main tools of cloning?
PCR, synthetic oligonucleotides, commercial vectors, various cloning methods,
How is it used?
For the overexpression of protein to be purified (for in vitro studies).
To study the effects of introducing or removing a gene from a cell or organism.
To study when and where a particular gene is turned on.
For creating genomic libraries that can be used for genomic sequencing.
For creating cDNA libraries that can be used for measuring transcription and
conducting function complementation experiments.
2. cDNA Library
What is a cDNA library? How are they made (in general terms)? How are they used?
3. Functional Complentation
What is functional complementation? How can it be used to determine the genetic
basis of a mutant phenotype in the lab?
What is the difference between forward and reverse genetics?
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