Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)and ion pair-reverse phase high performance liquid chromatography (IP-RP HPLC) techniques were combined to determine the sequence identity of short single-stranded deoxyoligonucleotides. In demonstrating this methodology a commercially abailable multiplex set of 8 primer pairs used for amplifying 8 specific loci, but with unknown sequences were separated and collected by IP-RP HPLC. Partial sequence information for IP-RP HPLC fractions was obtained from analyzing exonuclease digestion products by MALDI-TOF MS. By utilizing IP-RP HPLC, MALDI-TOF MS, exonuclease digests, and a simple computational algorithm an integrated strategy for determining the sequence of short necleic acid oligomers is demonstrated.
Citation: Analytical Biochemistry
Pub Type: Journals
informatics, MALDI-TOF MS, nucleic acid, PCR