“Cloning” as a molecular biology term refers to the manipulation of small DNA fragments to create DNA assemblies that can be used to express RNAs and protein coding genes in organisms. E. coli is often used because of the ease of manipulating DNA using this bacteria. Specifically, E. coli is simple to grow, grows rapidly, can reliably be transformed with DNA and is highly efficient at amplifying plasmid DNA allowing for rapid production of even milligram quantifies of DNA. The techniques used in DNA molecular biology are limited in number and relatively easy to perform. They include basic bacteriology to grow E. coli and basic biochemistry to manipulate DNA. Specifically, they include production of competent cells for introducing DNA into E. coli, isolation of plasmid DNA, restriction enzyme cleavage of DNA, gel electrophoresis, polymerase chain reaction amplification of DNA, and several methods for assembly of DNA fragments into plasmids. An overview of restriction enzymes and PCR is provided for those unfamilar with these techniques.