The enhancement of PCR amplification of a random sequence DNA library by DMSO and betaine: application to in vitro combinatorial selection of aptamers

J Biochem Biophys Methods. 2005 Aug 31;64(2):147-51. doi: 10.1016/j.jbbm.2005.06.003.

Abstract

A PCR method for uniform amplification of a random sequence DNA library is described. A combination of 1 M betaine and 5% DMSO improves the PCR amplification by increasing the ratio of full-length products to shortened products, which are a consequence of nonuniform amplification due to stable secondary structures in the templates. This method is expected to be beneficial for obtaining high-affinity aptamers with stable secondary structures.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Betaine / chemistry
  • Betaine / pharmacology*
  • DNA / chemistry
  • DNA / genetics*
  • Dimethyl Sulfoxide / chemistry
  • Dimethyl Sulfoxide / pharmacology*
  • Gene Library*
  • Nucleic Acid Conformation
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Solvents / chemistry
  • Solvents / pharmacology
  • Temperature

Substances

  • Solvents
  • Betaine
  • DNA
  • Dimethyl Sulfoxide