Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (95K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (50)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Kirpekar, F.
Right arrow Articles by Hillenkamp, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kirpekar, F.
Right arrow Articles by Hillenkamp, F.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, Vol 26, Issue 11 2554-2559, Copyright © 1998 by Oxford University Press

ARTICLES

DNA sequence analysis by MALDI mass spectrometry

F Kirpekar, E Nordhoff, LK Larsen, K Kristiansen, P Roepstorff and F Hillenkamp
Department of Molecular Biology, Odense University, Campusvej 55, DK- 5230 Odense M, Denmark. f.kir@PR-group.ou.dk

Conventional DNA sequencing is based on gel electrophoretic separation of the sequencing products. Gel casting and electrophoresis are the time limiting steps, and the gel separation is occasionally imperfect due to aberrant mobility of certain fragments, leading to erroneous sequence determination. Furthermore, illegitimately terminated products frequently cannot be distinguished from correctly terminated ones, a phenomenon that also obscures data interpretation. In the present work the use of MALDI mass spectrometry for sequencing of DNA amplified from clinical samples is implemented. The unambiguous and fast identification of deletions and substitutions in DNA amplified from heterozygous carriers realistically suggest MALDI mass spectrometry as a future alternative to conventional sequencing procedures for high throughput screening for mutations. Unique features of the method are demonstrated by sequencing a DNA fragment that could not be sequenced conventionally because of gel electrophoretic band compression and the presence of multiple non-specific termination products. Taking advantage of the accurate mass information provided by MALDI mass spectrometry, the sequence was deduced, and the nature of the non- specific termination could be determined. The method described here increases the fidelity in DNA sequencing, is fast, compatible with standard DNA sequencing procedures, and amenable to automation.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
B. Spottke, J. Gross, H.-J. Galla, and F. Hillenkamp
Reverse Sanger sequencing of RNA by MALDI-TOF mass spectrometry after solid phase purification
Nucleic Acids Res., July 7, 2004; 32(12): e97 - e97.
[Abstract] [Full Text] [PDF]

Home page
 Genome ResHome page
H. Ruparel, M. E. Ulz, S. Kim, and J. Ju
Digital Detection of Genetic Mutations Using SPC-Sequencing
Genome Res., February 1, 2004; 14(2): 296 - 300.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
C. T. Madsen, J. Mengel-Jorgensen, F. Kirpekar, and S. Douthwaite
Identifying the methyltransferases for m5U747 and m5U1939 in 23S rRNA using MALDI mass spectrometry
Nucleic Acids Res., August 15, 2003; 31(16): 4738 - 4746.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. Kaetzke and K. Eschrich
Simultaneous determination of different DNA sequences by mass spectrometric evaluation of Sanger sequencing reactions
Nucleic Acids Res., November 1, 2002; 30(21): e117 - e117.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. R. Edwards, Y. Itagaki, and J. Ju
DNA sequencing using biotinylated dideoxynucleotides and mass spectrometry
Nucleic Acids Res., November 1, 2001; 29(21): e104 - e104.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Shahgholi, B. A. Garcia, N. H. L. Chiu, P. J. Heaney, and K. Tang
Sugar additives for MALDI matrices improve signal allowing the smallest nucleotide change (A:T) in a DNA sequence to be resolved
Nucleic Acids Res., October 1, 2001; 29(19): e91 - e91.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y.-S. Kwon, K. Tang, C. R. Cantor, H. Koster, and C. Kang
DNA sequencing and genotyping by transcriptional synthesis of chain-terminated RNA ladders and MALDI-TOF mass spectrometry
Nucleic Acids Res., February 1, 2001; 29(3): e11 - e11.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
E. Nordhoff, C. Luebbert, G. Thiele, V. Heiser, and H. Lehrach
Rapid determination of short DNA sequences by the use of MALDI-MS
Nucleic Acids Res., October 15, 2000; 28(20): e86 - e86.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
A. RAY and B. NORDÉN
Peptide nucleic acid (PNA): its medical and biotechnical applications and promise for the future
FASEB J, June 1, 2000; 14(9): 1041 - 1060.
[Abstract] [Full Text]

Home page
 Nucleic Acids ResHome page
F. Hausch and A. Jaschke
Multifunctional DNA conjugates for the in vitro selection of new catalysts
Nucleic Acids Res., April 15, 2000; 28(8): e35 - e.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
A. Harksen, P. M. Ueland, H. Refsum, and K. Meyer
Four Common Mutations of the Cystathionine {beta}-Synthase Gene Detected by Multiplex PCR and Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry
Clin. Chem., August 1, 1999; 45(8): 1157 - 1161.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.