Methods and systems for molecular library generation

Abstract

The present disclosure provides methods and systems for generation of libraries comprising two or more sets of molecules. Libraries may comprise sets of molecules of different types. Sets of molecules may be generated by attachment to supports. Different types of molecules may be generated combinatorially, thereby generating a large number of unique molecules. One or more types of molecules may be used to identify one or more additional types of molecules. Libraries of the present disclosure may be used in, for example, nucleic acid sequencing.

1 . A method for generating a plurality of molecules, comprising:
 (a) providing a plurality of supports; and   (b) combinatorially generating a plurality of molecules coupled to and/or comprised in each of said plurality of supports, wherein a support of said plurality of supports comprises a first set of molecules and a second set of molecules of said plurality of molecules, which first set of molecules and second set of molecules are of different types.   
     
     
         2 . The method of  claim 1 , wherein said first set of molecules comprises deoxyribonucleic acid (DNA) molecules and said second set of molecules comprises molecules selected from the group consisting of (i) ribonucleic acid (RNA) molecules, (ii) peptide molecules, (iii) chemical compounds each having a molecular weight less than 900 Daltons, and (iv) molecules that are not DNA and RNA. 
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , wherein said first set of molecules comprises ribonucleic acid (RNA) molecules and said second set of molecules comprises molecules selected from the group consisting of (i) peptide molecules, (ii) chemical compounds each having a molecular weight less than 900 Daltons, and (iii) molecules that are not DNA and RNA. 
     
     
         5 .- 7 . (canceled) 
     
     
         8 . The method of  claim 1 , wherein said first set of molecules comprises one or more nucleic acid barcode sequences. 
     
     
         9 . The method of  claim 1 , wherein said plurality of supports is a plurality of beads. 
     
     
         10 . The method of  claim 1 , wherein molecules of said first set of molecules have sequences that encode molecules of said second set of molecules. 
     
     
         11 . (canceled) 
     
     
         12 . The method of  claim 1 , wherein at least a subset of said plurality of molecules is combinatorially generated by sequentially coupling one or more molecules to said plurality of supports or sequentially capturing one or more molecules within said plurality of supports. 
     
     
         13 . The method of  claim 12 , wherein said at least said subset of said plurality of molecules comprises a first molecule comprising a first segment and a second molecule comprising a second segment, and wherein said combinatorially generating comprises coupling said first molecule to a molecule coupled to a support of said plurality of supports and coupling said second molecule to said first molecule. 
     
     
         14 .- 16 . (canceled) 
     
     
         17 . The method of  claim 13 , further comprising coupling a third molecule comprising a third segment to said second molecule, wherein said third segment is different from said first segment or said second segment. 
     
     
         18 .- 21 . (canceled) 
     
     
         22 . The method of  claim 12 , wherein said coupling comprises partitioning said at least said subset of said plurality of molecules in separate partitions, and coupling said one or more molecules to each of said plurality of supports in said separate partitions. 
     
     
         23 .- 25 . (canceled) 
     
     
         26 . The method of  claim 12 , wherein said plurality of supports is a plurality of beads, and wherein said capturing comprises encapsulating said one or more molecules in said plurality of beads. 
     
     
         27 . The method of  claim 12 , wherein said capturing comprises partitioning said at least said subset of said plurality of molecules in separate partitions, and capturing said one or more molecules in each of said plurality of supports in said separate partitions. 
     
     
         28 .- 29 . (canceled) 
     
     
         30 . A method for generating a plurality of molecules, comprising:
 (a) providing a plurality of supports in a first plurality of partitions comprising a first set of molecules and a second set of molecules, which first set of molecules and second set of molecules are of different types;   (b) coupling said first set of molecules and said second set of molecules to said plurality of supports;   (c) providing said plurality of supports in a second plurality of partitions comprising a third set of molecules and a fourth set of molecules which third set of molecules and fourth set of molecules are of different types; and   (d) coupling said third set of molecules and said fourth set of molecules to said plurality of supports.   
     
     
         31 . The method of  claim 30 , wherein (i) said first set of molecules and said third set of molecules comprise deoxyribonucleic acid (DNA) molecules and (ii) said second set of molecules and said fourth set of molecules comprise ribonucleic acid (RNA) molecules. 
     
     
         32 . The method of  claim 30 , wherein (i) said first set of molecules and said third set of molecules comprise deoxyribonucleic acid (DNA) molecules and (ii) said second set of molecules and said fourth set of molecules comprise molecules selected from the group consisting of (A) peptide molecules, (B) chemical compounds each having a molecular weight less than 900 Daltons, and (C) molecules that are not DNA and RNA. 
     
     
         33 . The method of  claim 30 , wherein (i) said first set of molecules and said third set of molecules comprise ribonucleic acid (RNA) molecules and (ii) said second set of molecules and said fourth set of molecules comprise molecules selected from the group consisting of (A) peptide molecules, (B) chemical compounds each having a molecular weight less than 900 Daltons, and (C) molecules that are not DNA and RNA. 
     
     
         34 .- 36 . (canceled) 
     
     
         37 . The method of  claim 30 , wherein (d) comprises coupling said third set of molecules to said first set of molecules or to said plurality of supports. 
     
     
         38 . (canceled) 
     
     
         39 . The method of  claim 30 , wherein (d) comprises coupling said fourth set of molecules to said second set of molecules or to said plurality of supports. 
     
     
         40 . (canceled) 
     
     
         41 . The method of  claim 30 , wherein said first set of molecules and/or said third set of molecules comprises one or more nucleic acid barcode sequences. 
     
     
         42 . The method of  claim 30 , wherein said plurality of supports is a plurality of beads. 
     
     
         43 . The method of  claim 30 , wherein said first set of molecules and said third set of molecules positively identify said second set of molecules and said fourth set of molecules. 
     
     
         44 . The method of  claim 30 , further comprising (e) providing said plurality of supports in a third plurality of partitions comprising a fifth set of molecules and a sixth set of molecules which fifth set of molecules and sixth second set of molecules are of different types; and (f) coupling said fifth set of molecules and said sixth set of molecules to said plurality of supports. 
     
     
         45 . (canceled) 
     
     
         46 . The method of  claim 30 , wherein said first plurality of partitions is a plurality of wells or a plurality of droplets. 
     
     
         47 . (canceled) 
     
     
         48 . The method of  claim 30 , wherein said second plurality of partitions is a plurality of wells or a plurality of droplets. 
     
     
         49 .- 53 . (canceled)