Improving spot homogeneity by using polymer substrates in matrix-assisted laser desorption/ionization mass spectrometry of oligonucleotides.
We describe a method for improving the homogeneity of MALDI samples prepared for analysis of small, single-stranded oligonucleotides using the widely used DNA matrix system, 3-hydroxypicolinic acid/picolinic acid/ ammonium citrate. This matrix system typically produces large crystals around the rim of the dried sample and requires tedious searching of this rim with the laser. However, when a substrate is prepared using both Nafion and a hydrophilic, high-molecular-weight polymer, such as linear polyacrylamide, linear poly(ethylene oxide), or methyl cellulose, oligonucleotide-doped matrix crystals tend to be smaller and more uniformly distributed across the entire spot, thus decreasing the time that is required for locating a usable signal. In addition to MALDI characterization of the spatial distribution of "sweet spots," fluorescence microscopy allows for imaging dye-labeled DNA in dried MALDI spots. The mechanism of enhanced uniformity may involve increased viscosity in the MALDI sample droplet due to partial solubilization of the substrate by the MALDI sample solvent as well as partitioning of the matrix or DNA between the solvent and the undissolved portion of the polymer substrate.
Kim
Y
Y
Chemical and Analytical Sciences Division, Oak Ridge National Laboratory, Tennessee 37831-6365, USA.
Hurst
G B
GB
Doktycz
M J
MJ
Buchanan
M V
MV
D001483:Base Sequence; D008854:Microscopy, Electron; D009841:Oligonucleotides / Q000032:analysis*; D011108:Polymers; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009841:Oligonucleotides; D011108:Polymers