Genotyping complex short tandem repeats using electrospray ionization Fourier transform ion cyclotron resonance multistage mass spectrometry

摘要

Abstract: Electrospray ionization Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry is a rapidly emerging, universal platform with the ability to provide detailed information regarding genetic variation and the up- and down regulation of their cognate gene products. Herein, we report our progress towards the development of ESI-FTICR mass spectrometry for the characterization of genomic regions which contain both a length and sequence polymorphism (i.e., complex short tandem repeats). Specifically, it is demonstrated for the first time that a high-quality ESI-FTICR mass spectrum of a 82-bp double- stranded PCR product derived from a single, 50 $mu@L PCR reaction with less than 10 $MUL 10$+$MIN@15$/ moles injected into the mass spectrometer can be routinely obtained. It is important to note that each measurement, which translates to an accurate genotype, is completed on the timescale of seconds. Progress towards the implementation of flow- injection analysis methodology to increase the throughput is also presented using an alternating injection of a 15-mer and 16-mer oligonucleotide.!100