MALDI-TOF mass spectrometric method for detection of hybridized DNA oligomers.
Two new approaches for nucleic acid hybridizations by MALDI-TOF mass spectrometry are described. Hybridization using genomic DNA without polymerase chain reaction was demonstrated. Total genomic DNA of bacteriophages bound to charge-modified nylon membranes was identified by the hybridization of species-specific oligonucleotide probes. lambda-Phage DNA and M13 were used for the test with good success. Since MALDI-TOF mass spectrometry can be used to measure the molecular weights of different probes, mass spectrometry can be used for the detection of hybridizations with multiple probes. We demonstrate that multiple-probe hybridization can be resolved by mass spectrometry. Six probes with different mass tag were used for hybridization on a single spot. MALDI-TOF mass spectrometry was successfully used to measure these probes simultaneously. This provides a simple nonradioactive method for multiplex hybridization analysis. It has the potential to drastically increase the speed for microarray hybridization analysis in the future.
Isola
N R
NR
Life Sciences Division, Oak Ridge National Laboratory, Tennessee 37831-6378, USA.
Allman
S L
SL
Golovlev
V V
VV
Chen
C H
CH
D017104:Bacteriophage M13 / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics*; D004279:DNA, Viral / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D004279:DNA, Viral; D015345:Oligonucleotide Probes