RT Journal
A1 Stanssens, Patrick
A1 Zabeau, Marc
A1 Meersseman, Geert
A1 Remes, Gwen
A1 Gansemans, Yannick
A1 Storm, Niels
A1 Hartmer, Ralf
A1 Honisch, Christiane
A1 Rodi, Charles P.
A1 Böcker, Sebastian
A1 van den Boom, Dirk
T1 High-Throughput MALDI-TOF Discovery of Genomic Sequence Polymorphisms
JF Genome Research 
JO Genome Research 
YR 2004 
FD January 01 
VO 14 
IS 1 
SP 126 
OP 133 
DO 10.1101/gr.1692304 
UL http://genome.cshlp.org/content/14/1/126.abstract 
AB We describe a comparative sequencing strategy that is based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analyses of complete base-specific cleavage reactions of a target sequence. The target is converted to a DNA/RNA mosaic structure after PCR amplification using in vitro transcription. Cleavage with defined specificity is achieved by ribonucleases. The set of cleavage products is subjected to mass spectrometry without prior fractionation. The presented resequencing assay is particularly useful for single-nucleotide polymorphism (SNP) discovery. The combination of mass spectra from four complementary cleavage reactions detects approximately 98% of all possible homozygous and heterozygous SNPs in target sequences with a length of up to 500 bases. In general, both the identity and location of the sequence variation are determined. This was exemplified by the discovery of SNPs in the human gene coding for the cholesteryl ester transfer protein using a panel of 96 genomic DNAs.