Minimal primer and primer-free SELEX protocols for selection of aptamers from random DNA libraries

Weihua Pan, Ping Xin, Gary A. Clawson

Research output: Contribution to journalArticlepeer-review

39 Scopus citations


Standard systematic evolution of ligands by exponential enrichment (SELEX) protocols require libraries that contain two primers, one on each side of a central random domain, which allow amplification of target-bound sequences via PCR or RT-PCR. However, these primer sequences cause nonspecific binding by their nature (generally adding about 20 nt on each end of the random sequence of about 30-40 nt), and can result in large numbers of false-positive binding sequences and/or interfere with good binding random sequences. Here, we have developed two DNA-based methods that reduce and/or eliminate the primer sequences from the target-binding step, thus reducing or eliminating the interference caused by the primer sequences. In these methods, the starting selection libraries contain a central random sequence that is: (i) flanked by only 2 nt on each side (minimal primer); or (ii) flanked only by either a 2- or 0-nt overhand on the 3′ end (primer-free). These methods allow primer regeneration and re-elimination after and before selection, are fast and simple, and don't require any chemical modifications for selection in a variety of conditions. Further, the selection rounds are performed with DNA oligomers, which are generally employed as end product aptamers.

Original languageEnglish (US)
Pages (from-to)351-360
Number of pages10
Issue number3
StatePublished - Mar 2008

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • General Biochemistry, Genetics and Molecular Biology


Dive into the research topics of 'Minimal primer and primer-free SELEX protocols for selection of aptamers from random DNA libraries'. Together they form a unique fingerprint.

Cite this