Matrix-assisted laser desorption/ionization mass spectrometry of DNA using photocleavable biotin

被引:10
作者
Hahner, S
Olejnik, J
Lüdemann, HC
Krzymañska-Olejnik, E
Hillenkamp, F
Rothschild, KJ
机构
[1] Boston Univ, Dept Phys, Boston, MA 02215 USA
[2] AmberGen Inc, Boston, MA 02215 USA
[3] Univ Munster, Inst Med Phys & Biophys, D-4400 Munster, Germany
[4] Boston Univ, Mol Biophys Lab, Boston, MA 02215 USA
来源
BIOMOLECULAR ENGINEERING | 1999年 / 16卷 / 1-4期
关键词
matrix assisted laser desorption/ionization mass spectroscopy; 5 '-PC-biotin; oligonucleotides; UV-MALDI; IR-MALDI;
D O I
10.1016/S1050-3862(99)00049-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Oligonucleotides containing a photocleavable biotin (5'-PC-biotin) were analyzed by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) with wavelengths in the ultraviolet (UV) and infrared (IR) from solution and after capture on streptavidin-coated agarose or magnetic beads. The analysis was used to monitor the release of the oligonucleotides as a result of photochemical cleavage of the biotinylated linker. Near-UV pulses (UV-MALDI) led to predominant release of the photocleaved product. In contrast, only the uncleaved analyte was detected using IR pulses (IR-MALDI). Results from MALDI analysis are also presented for DNA containing a photocleavable 5'-amino group which can be covalently linked to a variety of activated surfaces and marker molecules. In a demonstration of this approach, a 5'-PC-biotinylated 49 nt RNA oligonucleotide was enzymatically synthesized using a PC-biotin-r(AG) dinucleotide primer, captured on streptavidin coated magnetic beads and analyzed by UV-MALDI. Potential applications of photocleavable linkers combined with MALDI for the analysis of nucleic acids are discussed. (C) 1999 Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:127 / 133
页数:7
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