Figure 1.
Schematic outline of the general SELEX procedure.
Table 1.
SELEX protocol.
Figure 2.
Remelting curves of diversity assay.
a) diversity standard and b) selection rounds (1–10).
Table 2.
List of oligonucleotides identified by Illumina sequencing after 10 selection rounds and analyzed in this study with parts corresponding to the putative consensus sequences printed in bold.
Figure 3.
Microtiter plate binding assay (FLAA) of selection pools to immobilized streptavidin.
Depicted are rounds 1 to 10 with the original library pool indicated as round 0.
Figure 4.
Microtiter plate binding assay (FLAA) of single clones applied as synthetic oligonucleotides to immobilized streptavidin.
Figure 5.
SPR sensorgram of streptavidin-aptamer binding.
Streptavidin was covalently immobilized on a CM5-chip. Aptamer clone R10#17 with highest affinity determined by FLAA was injected at a concentrations ranging between 0.1 and 2 µM in binding buffer.
Figure 6.
SPR sensorgram of selected oligonucleotides for direct comparison of binding affinity at identical concentration (1 µM).
Figure 7.
Relative frequency of clones in the pools (top100) of round 1 to 10 as evaluated by next generation sequencing.
Figure 8.
Loss of background binders characterized as unique clones in sequenced pools over the entire selection procedure.
Figure 9.
Sequence motifs and distribution.
a) Motifs identified by MEME from some selection rounds. b) Frequency of simplified palindrome in the sequence pools (top100) of round 1 to 10.
Figure 10.
Heat map indicating mutation frequency in selected abundant clones over selection rounds 5 to10 listed in successive rows.
Red color represents high mutation rates in the respective positions whereas little ore no mutations are colored blue.