使用含尿素的流动相进行变性反相高效液相色谱法分析寡脱氧核苷酸。
Denaturing reversed-phase HPLC using a mobile phase containing urea for oligodeoxynucleotide analysis.
作者信息
Fueangfung Suntara, Yuan Yinan, Fang Shiyue
机构信息
a Department of Chemistry , Michigan Technological University , Houghton , Michigan , USA.
出版信息
Nucleosides Nucleotides Nucleic Acids. 2014;33(7):481-8. doi: 10.1080/15257770.2014.891740.
Denaturing reversed-phase (RP) high performance liquid chromatography (HPLC) is usually achieved by elevating column temperature. In this article, an alternative method involving using a mobile phase that contains urea and performing HPLC at room temperature is described. The efficacy of the new method was demonstrated by analyzing a 61-mer oligodeoxynucleotide (ODN) and double-stranded (ds) ODNs. The multiple peaks of the 61-mer ODN under normal conditions merged into one under the denaturing conditions. The broad single peaks of dsODNs under normal conditions were split into two sharp peaks.
变性反相（RP）高效液相色谱法（HPLC）通常通过升高柱温来实现。在本文中，描述了一种替代方法，即使用含有尿素的流动相并在室温下进行HPLC。通过分析一个61聚体寡脱氧核苷酸（ODN）和双链（ds）ODN证明了该新方法的有效性。正常条件下61聚体ODN的多个峰在变性条件下合并为一个峰。正常条件下dsODN的宽单峰分裂为两个尖锐峰。
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