A calibrated diversity assay for nucleic acid libraries using DiStRO--a Diversity Standard of Random Oligonucleotides

Nucleic Acids Res. 2010 Mar;38(4):e23. doi: 10.1093/nar/gkp1108. Epub 2009 Dec 3.

Abstract

We have determined diversities exceeding 10(12) different sequences in an annealing and melting assay using synthetic randomized oligonucleotides as a standard. For such high diversities, the annealing kinetics differ from those observed for low diversities, favouring the remelting curve after annealing as the best indicator of complexity. Direct comparisons of nucleic acid pools obtained from an aptamer selection demonstrate that even highly complex populations can be evaluated by using DiStRO, without the need of complicated calculations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calibration
  • DNA / standards
  • Gene Library*
  • Kinetics
  • Nucleic Acid Denaturation
  • Oligodeoxyribonucleotides / chemical synthesis
  • Oligodeoxyribonucleotides / standards*
  • Reference Standards
  • SELEX Aptamer Technique
  • Temperature

Substances

  • Oligodeoxyribonucleotides
  • DNA