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Figure 2 | BMC Biotechnology

Figure 2

From: One-step selection of Vaccinia virus-binding DNA aptamers by MonoLEX

Figure 2

Identification of high-affinity-binding aptamers by MonoLEX. A combinatorial DNA library (64 b in length) was applied to an affinity capillary column coated with complete heat-inactivated VACV. Bound aptamers were desorbed from cut column slices and amplified by real-time PCR. (a) Data derived from different segments along the affinity column show a cumulation of aptamers in distinct segments while other segments do not amplify bound aptamers. Color labels indicate segments which were used for further evaluation of the aptamer pools. Figure 2b to 2d show the change of fluorescence per change of temperature plotted versus the temperature. (b) Melting temperature analysis of polyclonal aptamer pools after PCR amplification showing aptamer pools with different nucleic acid composition. In two of the pools (A38 and A77) more than one melting temperature maximum was observed, indicating different aptamer sub-pools. (c) Melting temperature analysis after aptamer dot blotting with the target molecule and repeated amplification. In A38 and A77 only one of the two sub-pools was amplified, indicating that one aptamer is more efficiently amplified after binding. The inset shows the preceding amplification results of an aptamer blotting assay with VACV (red solid line) and a negative control (black dotted line). The relative fluorescence is plotted vs. the cycle number. (d) Melting profile of the chemically synthesized and further characterized A38 with its predicted two-dimensional structure.

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