Comparison of approaches for purifying and desalting polymerase chain reaction products prior to electrospray ionization mass spectrometry

Anal Biochem. 2010 Mar 15;398(2):272-4. doi: 10.1016/j.ab.2009.10.026. Epub 2009 Oct 20.

Abstract

We have compared the recovery and desalting efficiency of different methods (ethanol precipitation, dialysis/ultrafiltration, purification with commercial kits, anion-exchange chromatography) of purifying and desalting PCR products prior to analysis by high-resolution/high mass accuracy electrospray ionization mass spectrometry (LTQ-Orbitrap). The results support the use of anion-exchange chromatography, which shows excellent desalting efficiency with almost no adducts observed, along with a recovery of approximately 70% and the ability to purify approximately 10 samples in 45 min.

Publication types

  • Comparative Study

MeSH terms

  • Analytic Sample Preparation Methods / methods*
  • Chemical Precipitation
  • Chromatography, Ion Exchange
  • Ethanol / chemistry
  • Oligonucleotides / chemistry*
  • Oligonucleotides / genetics
  • Oligonucleotides / isolation & purification*
  • Polymerase Chain Reaction*
  • Salts / chemistry*
  • Spectrometry, Mass, Electrospray Ionization*
  • Time Factors
  • Ultrafiltration

Substances

  • Oligonucleotides
  • Salts
  • Ethanol