Quantitation of DNA-Encoded Libraries by qPCR.

Christopher S Kollmann

Methods in molecular biology (Clifton, N.J.) 2022

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DNA-encoded library (DEL) yields can be easily measured throughout the selection process using the quantitative polymerase chain reaction (qPCR) (Sannino A, Gabriele E, Bigatti M, Mulatto S, Piazzi J, Scheuermann J, Neri D, Donckele EJ, Samain F, Chembiochem Eur J Chem Biol 20:955-962, 2019). Samples taken throughout the selection process are diluted prior to amplification and compared to standards of known DNA concentration. Here, I describe a general protocol using a double-stranded DNA binding dye for reaction monitoring. This allows the selection process to be assessed at each step prior to preparation for sequencing. The same method has additional applications in the practice of DEL technology. © 2022. Springer Science+Business Media, LLC, part of Springer Nature.