Preparation of Chemically Modified DNA Library for SELEX via Incorporation of CLB-dUTP in Primer Extension Reaction.

Aptamers are single-stranded DNA or RNA molecules which bind with high specificity to the molecular target for which they have been selected. Through a number of unnatural modifications, the diversity of interactions available for this class of molecules can be greatly expanded, potentially leading to better binding properties. Herein we describe a method to prepare an initial chemically modified DNA library for SELEX. It comprises the synthesis of a modified nucleotide in the CuAAC reaction and its purification by an adapted HPLC method. Subsequent stage is the incorporation of the prepared modified nucleotide into a DNA library in a primer extension reaction and a quick and easy method of its purification using an electroelution device. This allows the recovery of the resulting chemically modified ssDNA library with high efficiency to kick off the aptamer selection process.