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- Title
Genotyping by mass spectrometric analysis of short DNA fragments.
- Authors
Laken, Steven J.; Jackson, Peta E.; Kinzler, Kenneth W.; Vogelstein, Bert; Strickland, Paul T.; Groopman, John D.; Friesen, Marlin D.
- Abstract
A method has been developed to produce small DNA fragments from PCR products for analysis of defined DNA variations by mass spectrometry. The genomic region to be analyzed is PCR-amplified with primers containing a sequence for the type IIS restriction endonuclease Bpml. Bpml digestion of the resultant PCR products yields fragments as small as seven bases, which are then analyzed by electro-spray ionization mass spectrometry. The approach was validated using seven different variants within the APC tumor suppressor gene, in which a perfect correlation was obtained with DNA sequencing. Both the sense and antisense strands were analyzed independently, and several variants can be analyzed simultaneously. These results provide the basis for a generally applicable and highly accurate method that directly queries the mass of variant DNA sequences.
- Subjects
DNA; GENOMICS
- Publication
Nature Biotechnology, 1998, Vol 16, Issue 13, p1352
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/4333