Systematic Evolution of Ligands by Exponential Enrichment / High-Throughput Systematic Evolution of Ligands by Exponential Enrichment

From the time that the first SELEX experiments were described by 3 independent groups in 1990 (Ellington et al., 1990) (Tuerk et al., 1990) (Sullenger et al., 1990), the method has been adapted to a wide range of technologies (Chen et al., 2016) (Takahashi et al., 2016). A highly multiplexed, parallel HT-SELEX method was developed for NGS (Jolma et al., 2010). A variation of SELEX-seq (Slattery et al., 2011) uses Nextera adapter sequences for efficient library preparation (Zhang et al., 2016).

In this method, proteins are expressed as fusions with streptavidin-binding peptide (SBP), conjugated to Gaussia luciferase, in the pD40htSELEX expression vector. Each DNA ligand contains a 14 bp randomized region (14N), and a 5 bp barcode that uniquely identifies the individual SELEX sample. Partially nested primers are used in successive SELEX rounds. A double-stranded DNA mixture containing all possible 14 bp sequences is incubated with a DNA-binding protein immobilized into a well of a 96-well plate, resulting in binding of DNA to the protein. After washing and elution, the resulting population of more specific sequences is amplified by PCR and sequenced (Caroli et al., 2016)



  • Could contain sequence bias


Illumina Library prep and Array Kit Selector


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