During aptamer selection process (SELEX), whether aptamer-protein conjugate interfere during PCR cycle?

01 May 2021 1 9K Report

I'm trying to quantify aptamer (received from SELEX round) by PCR. As after elution we got protein-aptamer complex, is it necessary to remove that protein from aptamer-protein complex & use only aptamer for PCR cycles?

If yes, then what will be the procedure. Thank you in advance.

Vincent Paul Schmitz

The concentration and quantity of target protein should be many magnitudes lower than the SELEX library. The PCR cycle should have a ~5min - 90-95C step upon initialization, which will unbind any aptamers still attached to the target protein as well as denaturating the protein itself so that your sought after aptamers will not re-conjugate; as the structure of the target protein will have changed drastically.

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