Automated solid-phase extraction for purification of single nucleotide polymorphism genotyping products prior to matrix-assisted laser desorption/ionisation time-of-flight mass spectrometric analysis.
In this article, we describe the application of a novel micro elution solid-phase extraction method for purification of short stretches of DNA products of single nucleotide polymorphisms (SNPs) prior to MALDI mass spectrometry analysis. An important feature of our method is that the purification columns containing a copolymer of divinylbenzene and N-vinylpyrrolidone can be used several times thereby significantly reducing costs. We implemented this DNA purification technology into a fully automated procedure including molecular biology, MALDI sample preparation, automated mass spectrometric analysis and SNP allele calling by software. Due to the facile purification protocol, the methodology shown could furthermore be used for other applications where efficient medium to high-throughput purification of nucleic acids is required.
Sauer
Sascha
S
Department Lehrach, Max-Planck-Institute for Molecular Genetics, Ihnestrasse 73, 14195 Berlin-Dahlem, Germany. sauer@molgen.mpg.de
Kepper
Pamela
P
Smyra
Anett
A
Dahl
Andreas
A
Ferse
Falk-Thilo
FT
Lehrach
Hans
H
Reinhardt
Richard
R
D000469:Alkaline Phosphatase / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000235:genetics; D017931:DNA Primers; D005838:Genotype; D007202:Indicators and Reagents; D009841:Oligonucleotides / Q000032:analysis; D020641:Polymorphism, Single Nucleotide; D012313:RNA / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D017931:DNA Primers; D007202:Indicators and Reagents; D009841:Oligonucleotides; D012313:RNA; D004247:DNA; D000469:Alkaline Phosphatase