Comparison of approaches for purifying and desalting polymerase chain reaction products prior to electrospray ionization mass spectrometry.
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.
Commissariat à l'Energie Atomique, iBiTec-S, Service de Pharmacologie et d'Immunoanalyse, Gif-sur-Yvette, France.
Hélène
H
Manduzio
Armelle
A
Martelet
Eric
E
Ezan
François
F
Fenaille
D053000:Analytic Sample Preparation Methods / Q000379:methods*; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D000431:Ethanol / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012492:Salts / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors; D014462:Ultrafiltration
D009841:Oligonucleotides; D012492:Salts; D000431:Ethanol