Use of a nitrocellulose film substrate in matrix-assisted laser


ACS Legacy Archive. Cite this:Anal. Chem. 67, 19, 3482-3490. Note: In lieu of an abstract, this is the article's first page. Click to increase image s...
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Anal. Chem. 1995, 67,3482-3490

Use of a Nitrocellulose Film Substrate in Matrix-Assisted Laser Desorption/lonization Mass Spectrometry for DNA Mapping and Screening Yan=HuiLiu, Jian Bai, Xiaoli Liang, and David M. Lubman" Department of Chemistry, The University of Michigan, Ann A h r , Michigan 48109-1055

Patrick J. Venta College of Veterinary Medicine, Michigan State University, East Lansing, Michigan 48824- 1314

A nitrocellulose film is used as a substrate in matrixassisted laser desorptiodionization mass spectrometry (MALDI MS) studies of DNA. PCR products and DNA fragment digests obtained from biochemical procedures can be analyzedwith the use of a nitrocellulose substrate in MALDI MS whereas no signal is observed with the use of a stainless steel substrate. In this method, on-probe p d c a l i o n allows for effective elimination of the interfering effects of salts, buffers, and other contaminants that are usually present in DNA samples, which serve as important limitingfactors in the DNA molecular ion yield in the MALDI process. The use of the nitrocellulose film substrate also appears to improve the shot-to-shot and sample-to-samplereproducibility of the ion yield due to the more homogeneous coverage of maWanalyte over the sample surface. With the use of the nitrocellulose substrate, DNA fragments of up to 622 base pairs in complex mixcures provide mass spectra with minimal sample pdcation. Although only species corresponding to single-strandedDNA were detected, a mass calibration scheme was established allowing an accuracy of within one base pair for fragments of