Analysis of double-stranded polymerase chain reaction products from the Bacillus cereus group by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.
The analysis of polymerase chain reaction (PCR) products by electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR) has been achieved. Specifically, a 105 base-pair nucleotide portion of the ribosomal spacer region was amplified in two members of the B. cereus group (i.e. B. thuringiensis and B. cereus) using PCR. These amplified regions were then analyzed by gel electrophoresis and ESI-FTICR. Based on the predicted sequence of the PCR products for each organism, the mass measurement using ESI-FTICR matched the theoretical mass within experimental error and was consistent with gel electrophoresis results. In contrast, for the typical several hour time-scale of the gel electrophoresis experiment, the mass spectrometric analysis was completed in a matter of minutes. To our knowledge, this constitutes the first report demonstrating the ionization and detection of a double-stranded PCR product by ESI-MS. This preliminary result indicates the potential use of ESI-MS to analyze PCR products on a rapid time-scale, with potential for medical and taxonomic applications.
Wunschel
D S
DS
Department of Microbiology and Immunology, University of South Carolina, School of Medicine, Columbia 29209, USA.
Fox
K F
KF
Fox
A
A
Bruce
J E
JE
Muddiman
D C
DC
Smith
R D
RD
D001409:Bacillus cereus / Q000737:chemistry* / Q000378:metabolism; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000378:metabolism; D017357:Cyclotrons; D004269:DNA, Bacterial / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005583:Fourier Analysis; D013058:Mass Spectrometry; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D012329:RNA, Bacterial / Q000032:analysis
D004269:DNA, Bacterial; D012329:RNA, Bacterial