Effective isolation of RNA aptamer through suppression of PCR bias.
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.
Tsuji
Shoutaro
S
Division of Cancer Therapy, Kanagawa Cancer Center Research Institute, Yokohama-shi, Kanagawa 241-0815, Japan. stsuji@gancen.asahi.yokohama.jp
Hirabayashi
Naomi
N
Kato
Shintaro
S
Akitomi
Joe
J
Egashira
Hazuki
H
Tanaka
Taku
T
Waga
Iwao
I
Ohtsu
Takashi
T
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
D052157:Aptamers, Nucleotide; D012313:RNA