Zagorevskii Dmitri V, Aldersley Michael F, Ferris James P
Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute,209C Cogswell Laboratory, Troy, New York 12180, USA.
J Am Soc Mass Spectrom. 2006 Sep;17(9):1265-70. doi: 10.1016/j.jasms.2006.05.012. Epub 2006 Jun 30.
Oligonucleotides synthesized on a montmorillonite catalyst were analyzed directly. By mixing the catalyst with a matrix (2,4,6-trihydroxyacetophenone or 6-aza-2-thiothymine) and dibasic ammonium citrate, higher molecular weight products were detected compared with "classical" methods such as gel electrophoresis and HPLC with UV as a detector. The oligomers (30-mers and higher) were detected by mass spectrometry even though their concentration was less than 10(-4)% of the total content of the RNA. This method is different from the (MALDI) analysis of the eluates from montmorillonite, which otherwise requires desalting. Placing reaction mixtures with a high concentration of buffers on homoionic, preferably Li-containing, montmorillonite does not require desalting.
对在蒙脱石催化剂上合成的寡核苷酸进行了直接分析。通过将催化剂与基质(2,4,6-三羟基苯乙酮或6-氮杂-2-硫胸腺嘧啶)和柠檬酸氢二铵混合,与诸如凝胶电泳和以紫外光为检测器的高效液相色谱等“经典”方法相比,检测到了更高分子量的产物。即使寡聚物(30聚体及以上)的浓度低于RNA总含量的10(-4)%,也能通过质谱检测到。该方法不同于对蒙脱石洗脱液的(基质辅助激光解吸电离)分析,后者否则需要脱盐。将含有高浓度缓冲液的反应混合物置于同离子的、最好是含锂的蒙脱石上,不需要脱盐。