Isola N R, Allman S L, Golovlev V V, Chen C H
Life Sciences Division, Oak Ridge National Laboratory, Tennessee 37831-6378, USA.
Anal Chem. 2001 May 1;73(9):2126-31. doi: 10.1021/ac0013711.
Two new approaches for nucleic acid hybridizations by MALDI-TOF mass spectrometry are described. Hybridization using genomic DNA without polymerase chain reaction was demonstrated. Total genomic DNA of bacteriophages bound to charge-modified nylon membranes was identified by the hybridization of species-specific oligonucleotide probes. lambda-Phage DNA and M13 were used for the test with good success. Since MALDI-TOF mass spectrometry can be used to measure the molecular weights of different probes, mass spectrometry can be used for the detection of hybridizations with multiple probes. We demonstrate that multiple-probe hybridization can be resolved by mass spectrometry. Six probes with different mass tag were used for hybridization on a single spot. MALDI-TOF mass spectrometry was successfully used to measure these probes simultaneously. This provides a simple nonradioactive method for multiplex hybridization analysis. It has the potential to drastically increase the speed for microarray hybridization analysis in the future.
描述了两种通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）进行核酸杂交的新方法。展示了无需聚合酶链反应使用基因组DNA的杂交。通过物种特异性寡核苷酸探针的杂交鉴定了结合到电荷修饰尼龙膜上的噬菌体的总基因组DNA。使用λ噬菌体DNA和M13进行测试，取得了良好的成功。由于MALDI-TOF-MS可用于测量不同探针的分子量，质谱可用于检测与多种探针的杂交。我们证明了多探针杂交可以通过质谱进行解析。使用六个具有不同质量标签的探针在单个点上进行杂交。MALDI-TOF-MS成功地用于同时测量这些探针。这为多重杂交分析提供了一种简单的非放射性方法。它有可能在未来大幅提高微阵列杂交分析的速度。
