title: Effective isolation of RNA aptamer through suppression of PCR bias.
abstract: An aptamer is a short RNA or DNA molecule that binds to a specific target. The main strategy for obtaining aptamers is systematic evolution of ligands by exponential enrichment (SELEX). Although various SELEX techniques have been devised and refined on the basis of the selection technique used, in most cases, the isolation of an aptamer still requires several trials or the use of special equipment. In the present study, we attempted SELEX in which PCR bias was suppressed by using RNA transcription to amplify nucleic acids. This procedure, which can be accomplished easily and inexpensively without special equipment, effectively simplifies the SELEX process. Using this SELEX, we obtained large numbers of RNA aptamers against the target that could not be isolated by standard SELEX. The results of our study suggest that exclusion of PCR bias may be far more important than previously assumed for isolating RNA aptamers via SELEX.
authors.affiliation: Division of Cancer Therapy, Kanagawa Cancer Center Research Institute, Yokohama-shi, Kanagawa 241-0815, Japan. stsuji@gancen.asahi.yokohama.jp
authors.forename: Shoutaro
authors.initials: S
authors.lastname: Tsuji
authors.forename: Naomi
authors.initials: N
authors.lastname: Hirabayashi
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kato
authors.forename: Joe
authors.initials: J
authors.lastname: Akitomi
authors.forename: Hazuki
authors.initials: H
authors.lastname: Egashira
authors.forename: Taku
authors.initials: T
authors.lastname: Tanaka
authors.forename: Iwao
authors.initials: I
authors.lastname: Waga
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohtsu
mesh_terms: D052157:Aptamers, Nucleotide / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide; D012313:RNA