Primer-free aptamer selection using a random DNA library.
Aptamers are high-affinity oligonucleotides which can be selected from large random libraries by systematic evolution of ligands by exponential enrichment (SELEX) protocols, with affinities and specificities comparable or better than antibodies. The SELEX protocols comprise multiple rounds of selection, each of which require regeneration of bound ligands, which in turn require fixed primer sequences flanking the random library regions. These fixed primer sequences can interfere (with false positives and negatives) with the selection process. Here we present a primer-free protocol using a random DNA library.
Pan
Weihua
W
Gittlen Cancer Research Foundation, Hershey Medical Center, Hershey, PA, USA.
Clawson
Gary A
GA
D002448:Cell Adhesion; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D017931:DNA Primers / Q000378:metabolism*; D015723:Gene Library; D006801:Humans; D009414:Nerve Growth Factors / Q000378:metabolism; D016133:Polymerase Chain Reaction; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000378:metabolism; D052156:SELEX Aptamer Technique / Q000379:methods*; D017422:Sequence Analysis, DNA; D013696:Temperature; D014158:Transcription, Genetic
D017931:DNA Primers; D009414:Nerve Growth Factors; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D004247:DNA