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Matrix‐assisted laser desorption/ionization time‐of‐flight analysis of low‐concentration oligonucleotides and mini‐sequencing products
Author(s) -
Liu Yiding,
Sun Xiyuan,
Guo Baochuan
Publication year - 2003
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1200
Subject(s) - chemistry , oligonucleotide , chromatography , desorption , reproducibility , matrix (chemical analysis) , matrix assisted laser desorption/ionization , sample preparation , analytical chemistry (journal) , dna , organic chemistry , adsorption , biochemistry
A new sample preparation procedure has been developed to improve the matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) analysis of low‐concentration oligonucleotides. In this method, a few μL of the oligonucleotide solutions are first dispensed and allow drying and shrinking to a small spot on an anchoring target. Thereafter, a small volume (0.1 μL) of the saturated 3‐hydroxypicolinic acid (3‐HPA) solution is added to the top of the shrunk oligonucleotide spot. It is found that the sample prepared by this procedure is more homogenous and substantially reduces the need to search for ‘sweet’ spots. Importantly, the increased shot‐to‐shot and sample‐to‐sample reproducibility makes it possible to perform high‐throughput and quantitative analysis. We demonstrated that this procedure allowed the robust detection of oligonucleotides at a level of 0.01 μM and mini‐sequencing products produced using only 50 fmol of the extension primer. Copyright © 2003 John Wiley & Sons, Ltd.