| 主权项 |
1. A method for producing a focused library comprising a plurality of different molecules, wherein each molecule is linked to an identifier oligonucleotide comprising tags identifying a plurality of reactants that participated in formation of the molecule, the method comprising the steps of:
i) producing an initial library comprising a plurality of different initial molecules, wherein each initial molecule is linked to an identifier oligonucleotide comprising tags identifying a plurality of reactants that participated in formation of the initial molecule, the initial library having a higher diversity than the focused library, and wherein said initial library is produced by a split-and-mix method comprising the steps of: a. providing, in separate compartments, a plurality of nascent bifunctional complexes each comprising a chemical reaction site and a priming site, and reacting the nascent bifunctional complexes at their chemical reaction sites with one or more reactants; b. reacting the priming sites of the plurality of nascent bifunctional complexes with one or more tags, wherein the reaction between the priming site and the one or more tags is catalyzed by an enzyme; wherein steps (a) and (b) provide a plurality of intermediate bifunctional complexes having a modified chemical reaction site containing a structural entity formed from the chemical reaction site and the one or more reactants and a modified priming site containing the one or more tags; c. pooling the intermediate bifunctional complexes into a single compartment to provide a mixture, and splitting the mixture into separate compartments d. reacting each compartment containing a mixture of intermediate bifunctional complexes with one or more additional reactants at the modified reactant sites and one or more additional tags identifying the one or more additional reactants at the modified priming sites using the methods of steps (a) and (b); and e. optionally repeating steps (c) and (d) as many times as desired ii) subjecting said initial library to a partitioning step, wherein said partitioning step provides a partitioned library synthesized from some, but not all, of the plurality of reactants used in the synthesis of the initial library, and iii) identifying the reactants used in the synthesis of at least some of the molecules of the partitioned library, iv) producing a focused library having a lower diversity than the initial library, wherein said focused library is produced by a split-and-mix method comprising the steps of: a. providing at least some of the reactants used in the synthesis of the molecules of the partitioned library, b. providing either i) at least some, but not all, of the reactants used in the synthesis of the initial library but not used in the synthesis of the partitioned library, and/or ii) further reactants not used in the synthesis of the initial library, c. reacting the reactants provided in steps (iv)(a) and (iv)(b) in a split-and-mix method comprising the steps cited in (i)(a)-(i)(e); and d. producing the focused library comprising a plurality of different molecules. |
