Analysis of the degradation of oligonucleotide strands during the freezing/thawing processes using MALDI-MS
Analysis of the degradation of oligonucleotide strands during the freezing/thawing processes using MALDI-MS
复制标题
DOI：
10.1021/ac000225s
复制
发表时间：
2000-10-15
影响因子：
7.4
通讯作者：
Bentzley, CM
中科院分区：
文献类型：
--
作者：
Davis, DL;O'Brien, EP;Bentzley, CM
长度范围为5至35个单位的合成寡核苷酸链通常用作各种生物分析应用中的标准品、探针和模板。直到最近，他们的准备，储存和处理被认为是不重要的，但这项工作提供了相反的有价值的信息。通过反复冷冻/解冻短链，然后进行基质辅助激光解吸电离质谱（MALDI-MS）分析，研究样品制备过程中寡核苷酸链的系统降解。这里显示，寡核苷酸链的寿命取决于几个因素，包括碱基组成、溶液浓度和链长度以及解冻条件。我们的研究表明，链的稳健性是碱基依赖的：T-mer > A-mer > C-mer > G-mer。同样，链长度的增加增加了样品降解的趋势。另一个观察结果包括混合碱样品根据结构构象降解。所有这些观察结果均归因于样品在冷冻期间样品/溶剂分离期间发生降解。
Synthetic oligonucleotide strands ranging from 5 to 35 units in length are commonly used as standards, probes, and templates in various bioanalytical applications. Until recently, their preparation, storage, and handling were regarded as unimportant, but this work provides valuable information to the contrary. The systematic degradation of oligonucleotide strands during sample preparation is investigated by repeatedly freezing/thawing short strands followed by matrix-assisted laser desorption ionization mass spectrometric (MALDI-MS) analysis. It is shown here that the longevity of an oligonucleotide strand is dependent on several factors including base composition, solution concentrations, and strand length as well as thawing conditions. Several trends in strand robustness were established, Our studies reveal that the robustness of strands is base-dependent: T-mer > A-mer > C-mer > G-mer. Likewise, an increase in the length of the strands increases the tendency of a sample to degrade. Another observation included that samples of mixed bases degrade according to structural conformations. All of these observations are attributed to the fact that the samples undergo degradation during sample/solvent isolation during freezing.