Testing the Various Efficiencies of Oligonucleotide Synthesis for Use in the Gene Therapy Treatment of Cancer
Morehouse, Yasha A.
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In an effort to produce ever more efficient oligonuleotide primers for genetic research, a comparison was made between various types of synthesizing machinery, synthesis methods and purification methods. A standard sequence (M13F0R24, see App. A) was run on each of the four synthesizing machines. The oligonucleotide primer product was then analyzed in four separate areas: 1) efficiency of synthesis: this information was provided by the machine itself and was based on the coupling efficiency at each step. 2) yield of the primer: an optical density reading was used to calculate both the concentration and yield of the product. 3) percent of full length product: chromatography separates the sample according to size and allows visualization of the amount of desired length primer. 4) performance in a sequencing application: the primer was used to sequence of piece of DNA to determine its efficiency in research applications. Based on the results obtained in each of these areas, it was concluded that Beckman 1000 synthesizing ,instruments are superior to Pharmacia-Biotech in synthesizing oligonucleotide primers.