Application of DNA-binding polymers for preparation of DNA for analysis by matrix-assisted laser desorption/ionization mass spectrometry.
The susceptibility of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to the presence of salts in a sample, especially salts of alkali metals, requires careful and often tedious desalting procedures which complicate and slow the throughput of MS-based methods. A novel approach to sample preparation was developed based on the extraction of DNA out of solution onto a solid surface with an attached DNA-binding polymer, such as polyethyleneimine or polyvinylpyrrolidone. The observed binding is strong enough to sustain washing, and, as a result, desalting and concentration can be performed in a single fast step. After DNA has been immobilized on the surface and supernatant solution removed, subsequent addition of MALDI matrix releases material from the surface, which co-crystallizes with matrix. The mass spectrometric analysis is then performed directly from this support. Analysis of oligonucleotides and three-fold multiplexed SNP typing reactions performed by this method shows improved sensitivity and excellent resolution for various DNA fragments, together with high tolerance to various buffer components, such as alkali metals and surfactants. Simplicity and speed make it attractive for high-throughput sample preparation and analysis of oligonucleotide mixtures by MALDI-MS.
Smirnov
I P
IP
Applied Biosystems, 500 Old Connecticut Path, Framingham, MA 01701, USA. smirnoip@appliedbiosystems.com
Hall
L R
LR
Ross
P L
PL
Haff
L A
LA
D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification*; D006801:Humans; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011094:Polyethyleneimine; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011205:Povidone; D012492:Salts; D017422:Sequence Analysis, DNA / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D012492:Salts; D011094:Polyethyleneimine; D004247:DNA; D011205:Povidone