Krylov, Sergey N.Tsushima, Robert2016-09-132016-09-132013-09http://hdl.handle.net/10315/31955Aptamers are short oligonucleotide sequences that are capable of binding with high affinity and specificity to a wide variety of targets. The selection of aptamers from a random oligonucleotide library, by cell-SELEX, has lead to their in vivo application in both research and clinical settings. Cell-SELEX continues to be hindered by the non-specific binding of selection sequences to the complex cell surface as well as low specificity in the PCR amplification of DNA. In this work the cell-SELEX procedure was optimized to improve the efficiency of aptamer selection using three modifications: masking DNA characterization, non-SELEX selection and touchdown PCR. An established masking DNA model was tested against the MCF-7 and 4T1 cell lines and was shown to be capable of determining the masking DNA concentration needed for aptamer selection. The traditional SELEX approach was combined with a non-SELEX aptamer selection protocol to reduce the concentration and heterogeneity of sequences collected after the first round of selection thus leading to more accurate PCR amplification and a decrease in byproduct formation. Lastly, touchdown PCR was used to successfully eliminate the amplification of genomic DNA reducing the formation of genomic DNA byproducts.Author owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.Electronic Thesis or DissertationAptamerscell-SELEXDNAPCRnon-SELEX