Minisequencing, PinPoint Assay, PCR Products, Nucleotide, Apolipoprotein, Proto Oncogene, Mass Tagged, Human Genomic, Voyager,Maldi Tof Platform

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Home >> Biotechnology and Genomics >> Mass Spectrometry - An Essential Tool for Genome and Proteome Analysis >>PinPoint Assay for SNP Detection Using MALDI-TOF MS of PCR Products.

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Pin Point Assay For SNP Detection Using MALDI-TOF MS Of PCR Products.
One of the methods that was described as PinPoint assay is based on using a primer directly upstream of a known point mutation or SNP, so that in PCR, the primer is extended by a single ddNTP. In other cases, a primary may anneal several bases away from SNP and extended more than one bases before incorporating a specific ddNTP, which terminates the reaction. The PCR products are solid-phase purified and detected by /z value specific for the nucleotides added in the extension reaction. The technique is described as minisequencing, since it involves sequencing of a segment few bases long.

SNPs in several genes like p53 gene, apolipoproteins E gene and RET proto-oncogene have been identified using this approach. Multiplex SNP analysis has also been possible using MALDI-TOF MS, as shown for PCR amplification from BRCA1 exon 13 locus having five SNPs. This is facilitated by using mass-tagged minisequencing primers (extra non-complementary dT nucleotide at the 5’ end) for each SNP position, so that all the five primers could be extended in a single reaction and PCR products could be simultaneously analysed by MALDI-TOF Ms. Mass tagged ddNTPs have also been used to increase the mass difference between PCR products, so that this mass difference can be resolved by MALDI-TOF MS. In another study multiplex genotyping of 12 different SNPs (selected from human genomic SNPs) was possible using minisequencing. The PCR products in the above technique can be analysed by Voyager DE MALDI-TOF platform.

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Isotope Ratio Mass Spectrometry - IRMS Applications of Mass Spectrometry for Genome Proteome Analysis DNA Sequencing by DE-MALDI-TOF MS SNP Detection Using MS Pinpoint Assay for SNP Detection Using MALDI-TOF MS of PCR Products SNP Detection Using PNA Probes SNP Detection Using Invader Cleavage SNP Detection Involving Minisequencing Using Maldi on a Chip Technology Proteome Analysis Using Mass Spectrometer Peptide Sequencing Determination of Molecular Weights of Intact Proteins Protein Identification by Combining Mass Spectrometer With Database Search Searching With Peptide Mass Fingerprints Searching With Tandem Mass Spectrometric Data Searching for Protein Modifications Analysis of Postranslational Modifications