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 diagnose genetic diseases and identify pathogens through the analysis of DNA. As clinical applications for molecular diagnosis increase, rapid, reliable methods for determination of DNA size will be needed. Mass spectrometry offers the potential of analyzing amplified DNA quickly and reliably, without the need for gel-based separation and sample labeling steps that are conventionally employed. Both electrospray ionization and matrix-assisted laser desorption/ionization have been evaluated for the size analysis of DNA using both synthetic oligonucleotides and PCR-amplified samples corresponding to bases 1626 to 1701 of the cystic fibrosis transmembrane conductance regulator gene. Both technologies have been demonstrated to have mass range and sensitivity required for the analysis of PCR-amplified DNA in this size range using minimal sample preparation. Steps required to incorporate either ionization technique into a reliable analytical scheme for the rapid, routine analysis of DNA are outlined.
Doktycz
M J
MJ
Health Sciences Research Division, Oak Ridge National Laboratory, Tennessee 37831-6365, USA.
Hurst
G B
GB
Habibi-Goudarzi
S
S
McLuckey
S A
SA
Tang
K
K
Chen
C H
CH
Uziel
M
M
Jacobson
K B
KB
Woychik
R P
RP
Buchanan
M V
MV
D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004247:DNA