Gene Synthesis - Toward a Future World

University of Colorado Department of Chemistry Boulder, Colorado 

BamKL restriction endonuclease. The symbol p represents a 5 -phosphate. 

An example illustrating the insertion of a foreign DNA segment into the tetracycline gene of pBR322 at the BamWL site. Tet and Amp refer to tetracycline and ampicillin genes, respectively. 

There are two basic approaches for sequencing DNA. These methodologies have been published in considerable detail. 

The insoluble pol3nner support is synthesized from silica gel and an appropriately derivatized deoxynucleoside. We routinely start with high performance liquid chromatography (HPLC) grade silica gel as a support matrix.The initial synthesis step involves refluxing 3-aminopropyl triethoxysilane with silica gel in dry toluene for 3 h to form the silylamine. This derivatized silica gel is then allowed to react with an appropriately protected nucleoside containing a 3 -p-nitrophenyl-succinate ester. The product is the nucleoside covalently linked to the support (compounds la-d). 

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