The capability of tandem spectrometry to detect single and multiple SNPs in a single stretch of DNA will be evaluated. The p53 gene contains polymorphisms in codon 47 (TCG and CCG) and codon 47 (CGC and CCC). This will be studied as a model system. In initial work a 67 bp PCR product containing only the C to T switch will be analyzed by mass spectrometry (MS). In subsequent work 120 bp PCR products containing both alleles will be analyzed by MS2. This is pushing the envelope for mass discrimination using triple quadrupole or ion trap tandem mass spectrometry. The use of stable isotope labeled nucleotides increases the mass differences on C to T substitution (from 15 to 27 mass) units. This and the use of MSn (by sequential fragmentation of the molecule into smaller pieces) may be necessary to analyze larger PCR products.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
1F31HG002320-01
Application #
6312489
Study Section
Special Emphasis Panel (ZRG1-MDCN-7 (02))
Program Officer
Graham, Bettie
Project Start
2000-09-25
Project End
Budget Start
2000-09-25
Budget End
2001-09-24
Support Year
1
Fiscal Year
2000
Total Cost
$20,560
Indirect Cost
Name
University of South Carolina at Columbia
Department
Pathology
Type
Schools of Medicine
DUNS #
111310249
City
Columbia
State
SC
Country
United States
Zip Code
29208