Enzymatic encoding methods for efficient synthesis of large libraries

Abstract

Disclosed is a method for obtaining a bifunctional complex comprising a molecule linked to a single stranded identifier oligonucleotide, wherein a nascent bifunctional complex comprising a chemical reaction site and a priming site for enzymatic addition of a tag is a) reacted at the chemical reaction site with one or more reactants, and b) reacted enzymatically at the priming site with one or more tag(s) identifying the reactant(s).

1 - 161 . (canceled) 
     
     
         162 . A split-and-mix method for synthesising a library of different bifunctional complexes comprising a small molecule linked to an identifier oligonucleotide, the method comprising:
 i) dividing a plurality of nascent bifunctional complexes comprising a nascent small molecule linked to one or more oligonucleotide tags into multiple fractions;   ii) reacting a reactant with said nascent bifunctional molecule;   iii) chemically ligating a single stranded oligonucleotide tag identifying said reactant to an oligonucleotide tag of said nascent bifunctional molecule,   iv) providing one or more anti-tags capable of hybridizing to one or more chemically ligated oligonucleotide tags;   v) hybridizing said one or more anti-tags to said one or more chemically ligated oligonucleotide tags after each synthesis round or after a final round of synthesis;   vi) polymerase extending said anti-tags to generate complementary oligonucleotide tags; and   vii) enzymatically or chemically ligating said complementary oligonucleotide tags.