SSCP screening of individual aptamers

Biotechniques. 2001 Oct;31(4):828, 830, 832, 834. doi: 10.2144/01314st10.

Abstract

Aptamers are specific binding nucleic acids that emerge from in vitro selection. During the systematic evolution of ligands by exponential enrichment (SELEX) procedure, analysis of the sequences of the numerous selected individual molecules becomes an important step in the final stage of aptamer selection. The sequencing of cloned aptamers from the selected pool generally reveals groups of identical sequences and rarely occurring individual aptamers. This study demonstrates an approach similar to the single strand conformation polymorphism (SSCP) method used for mutation testing in genes. Human angiotensin I-specific aptamers have been used to show the efficiency of the SSCP method to classify selected individual sequences into identity groups, which minimizes sequencing efforts. Additionally this approach allows the rapid isolation and identification of aptamers from a mixture.

MeSH terms

  • Angiotensin I / genetics
  • Base Sequence
  • Biotechnology
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA, Single-Stranded / genetics*
  • DNA, Single-Stranded / isolation & purification*
  • Humans
  • Polymerase Chain Reaction / methods
  • Polymorphism, Single-Stranded Conformational*

Substances

  • DNA Primers
  • DNA, Single-Stranded
  • Angiotensin I