ANALYSIS OF POLYMERASE CHAIN REACTION-AMPLIFIED DNA PRODUCTS BY MASS-SPECTROMETRY USING MATRIX-ASSISTED LASER-DESORPTION AND ELECTROSPRAY -CURRENT STATUS

Recent advances in molecular biology are making it possible to diagnos e genetic diseases and identify pathogens through the analysis of DNA. As clinical applications for molecular diagnosis increase, rapid, rel iable methods for determination of DNA size will be needed. Mass spect rometry offers the potential of analyzing amplified DNA quickly and re liably, without the need for gel-based separation and sample labeling steps that are conventionally employed. Both electrospray ionization a nd matrix-assisted laser desorption/ionization have been evaluated for the size analysis of DNA using both synthetic oligonucleotides and PC R-amplified samples corresponding to bases 1626 to 1701 of the cystic fibrosis transmembrane conductance regulator gene. Both technologies h ave been demonstrated to have mass range and sensitivity required for the analysis of PCR-amplified DNA in this size range using minimal sam ple preparation. Steps required to incorporate either ionization techn ique into a reliable analytical scheme for the rapid, routine analysis of DNA are outlined. (C) 1995 Academic Press, Inc.