High-throughput MALDI-TOF discovery of genomic sequence polymorphisms.

摘要: 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 target sequence. The converted to DNA/RNA mosaic structure after PCR amplification using in vitro transcription. Cleavage with defined specificity achieved by ribonucleases. set products subjected without prior fractionation. presented resequencing assay particularly useful for single-nucleotide polymorphism (SNP) discovery. combination spectra from four complementary detects approximately 98% all possible homozygous and heterozygous SNPs sequences length up 500 bases. In general, both the identity location sequence variation are determined. This was exemplified discovery human gene coding cholesteryl ester transfer protein panel 96 genomic DNAs.