Back to Search
Start Over
MALDI-TOF Mass Spectrometric Detection of Multiplex Single Base Extended Primers. A Study of 17 Y-Chromosome Single-Nucleotide Polymorphisms.
- Source :
-
Analytical Chemistry . 10/15/2004, Vol. 76 Issue 20, p6039-6045. 7p. - Publication Year :
- 2004
-
Abstract
- One of the most promising techniques for typing of multiple single-nucleofide polymorphism (SNP) is detection of single base extension primers (SBE) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We present a new MALDITOF MS protocol for typing of multiple SNPs in a single reaction. Biotin-labeled ddNTPs were used in the SBE reaction and solid phase-bound monomeric avidin was used as capturing/purification scheme allowing the exclusive release of the SBE products under gentle condidons using 5% triethylantine. We dubbed this method monomeric avidin triethylamine purification. The biotinlabeled dd.NTPs contained linkers with different masses ensuring a clear separation of the alleles even for SBE primers with a mass of 10 300 Da. Furthermore, only 25-350 fmol of SBE primers were necessary in order to obtain reproducible MALDI-TOF spectra. Similar signal intensities were obtained in the 5500-10 300 m/z mass range by increasing the concentration of the longer SBE primers in the reaction. To validate the technique, 17 Y-chromosome SNPs were analyzed in 200 males. The precision and accuracy of the mass determination were analyzed by parametric statistic, and the potential use of MALDI-TOF MS for SNP typing is discussed. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00032700
- Volume :
- 76
- Issue :
- 20
- Database :
- Academic Search Index
- Journal :
- Analytical Chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 14893869
- Full Text :
- https://doi.org/10.1021/ac049264k