One of the major difficulties that arises when selecting aptamers containing a G-quadruplex is the correct amplification of the ssDNA sequence. Can aptamers containing a G-quadruplex be selected from a degenerate library using non-equilibrium capillary electrophoresis (CE) of equilibrium mixtures (NECEEM) along with high-throughput Illumina sequencing? In this article, we present some mismatches of the G-quadruplex T29 aptamer specific to thrombin, which was PCR amplified and sequenced by Illumina sequencing. Then, we show the proportionality between the number of sequenced molecules of T29 added to the library and the number of sequences obtained in Illumina sequencing, and we find that T29 sequences from this aptamer can be detected in a random library of ssDNA after the sample is fractionated by NECEEM, amplified by PCR, and sequenced. Treatment of the data by the counting of double-stranded DNA T29 sequences containing a maximum of two mismatches reveals a good correlation with the enrichment factor (f_E). This factor is the ratio of the number of aptamer sequences found in the collected complex sample divided by the total number of sequencing reads (aptamer and non-aptamer) plus the quantity of T29 molecules (spiked into a DNA library) injected into CE.

选择含有G-四链体的适体时出现的主要困难之一是正确扩增ssDNA序列。是否可以使用平衡混合物（NECEEM）的非平衡毛细管电泳（CE）和高通量Illumina测序从简并文库中选择含有G-四链体的适体？在本文中，我们介绍了凝血酶特异的G-四链体T29适体的一些错配，并通过Illumina测序对其进行了PCR扩增和测序。然后，我们显示了添加到该文库中的T29测序分子的数量与Illumina测序中获得的序列数量之间的比例，并且发现在样品被分离后，可以在ssDNA随机文库中检测到该适体的T29序列经NECEEM分离，PCR扩增并测序。f _E）。该因子是在收集的复杂样品中发现的适体序列数除以测序读数总数（适体和非适体）与注入CE的T29分子（掺入DNA文库）的数量之比。