A method of thermal dissociation of DNA–protein conjugates was developed for the selection of aptamers to human alpha-fetoprotein. The method is based on the identification of the DNA oligonucleotides from the combinatorial DNA libraries with the highest specificity to targets, which remained within the conjugate after two stages: a) washing of unbound sequences; and b) removal of the sequences bound to the DNA but displaying a low affinity to it. The undestroyed most stable complexes were subjected to the thermal dissociation with the involvement of the corresponding oligonucleotides in the next rounds of selection. A comparison of the proposed method with the standard washing demonstrated a more intensive enrichment of the library. Due to the tighter conditions for washing of the target-bound DNA oligonucleotides the use of thermal dissociation can increase the efficiency of selection of highly specific aptamers to their molecular targets.

中文翻译：

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使用热解离选择 DNA 适体

开发了一种 DNA-蛋白质偶联物的热解离方法，用于选择人类甲胎蛋白的适体。该方法基于从对靶标具有最高特异性的组合 DNA 文库中鉴定出的 DNA 寡核苷酸，这些寡核苷酸在两个阶段后仍保留在偶联物中： a) 洗涤未结合的序列；b) 去除与 DNA 结合但对其显示出低亲和力的序列。未破坏的最稳定的复合物经受热解离，其中相应的寡核苷酸参与下一轮选择。所提出的方法与标准洗涤的比较表明文库的富集程度更高。