Purification of single-stranded DNA by co-polymerization with acrylamide and electrophoresis

Tulsi Ram Damase, Andrew D Ellington, Peter B Allen

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Single-stranded DNA (ssDNA) oligonucleotides are useful as aptamers, hybridization probes and for emerging applications in DNA nanotechnology. Current methods to purify ssDNA require both a strand-separation step and a separate size-separation step but may still leave double-stranded DNA (dsDNA) impurities in the sample. Here, we use commercially available acrydite DNA primers to immobilize one strand of a PCR product within a polyacrylamide matrix. Electrophoresis moves the non-crosslinked DNA into the gel where the single-stranded product of desired size can be recovered. Our results show this method produces high yields of pure ssDNA.

Original languageEnglish (US)
Pages (from-to)275-282
Number of pages8
JournalBioTechniques
Volume62
Issue number6
DOIs
StatePublished - Jun 1 2017

Keywords

  • Acrylamide/chemistry
  • Aptamers, Nucleotide/chemistry
  • DNA Primers/chemistry
  • DNA, Single-Stranded/chemistry
  • Electrophoresis, Polyacrylamide Gel/instrumentation
  • Equipment Design
  • Immobilized Nucleic Acids/chemistry
  • Polymerization

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