Sample multiplexing

Abstract

The invention generally relates to methods for sample multiplexing. In certain embodiments, methods of the invention obtaining a plurality of different reactant molecules, attaching a unique identifier to the reactant molecules, and forming a droplet including the reactant molecules.

1 - 29 . (canceled) 
     
     
         30 . A sample preparation method, the method comprising:
 attaching an adaptor oligonucleotide to a nucleic acid, wherein the adaptor oligonucleotide comprises a universal primer binding site;   conducting an amplification reaction using a set of primers comprising a first primer species that hybridizes to the universal primer site of the adaptor oligonucleotide and a second primer species that hybridizes to a target site on the nucleic acid; and   conducting a second amplification reaction on products of the first amplification reaction to generate amplification products.   
     
     
         31 . The method of  claim 30 , wherein the adaptor oligonucleotide further comprises a barcode sequence. 
     
     
         32 . The method of  claim 30 , wherein the amplification reaction comprises a polymerase chain reaction. 
     
     
         33 . The method of  claim 30 , further comprising sequencing the amplification products. 
     
     
         34 . The method of  claim 30 , wherein sequencing comprises sequencing-by-synthesis 
     
     
         35 . The method of  claim 30 , wherein prior to sequencing, the amplification products are attached directly or indirectly to a solid support. 
     
     
         36 . The method according to  claim 30 , wherein attaching comprises ligating the adaptor oligonucleotide to the nucleic acid. 
     
     
         37 . The method according to  claim 30 , wherein prior to the attaching step, the nucleic acid is fragmented. 
     
     
         38 . The method according to  claim 37 , further comprising end-repairing the fragmented nucleic acid. 
     
     
         39 . The method according to  claim 30 , wherein the nucleic acid comprises DNA. 
     
     
         40 . The method according to  claim 30 , wherein the nucleic acid comprises RNA. 
     
     
         41 . The method according to  claim 40 , wherein prior to the attaching step, the method further comprises converting the RNA to cDNA. 
     
     
         42 . The method according to  claim 30 , wherein the second amplification reaction comprises a nested polymerase chain reaction. 
     
     
         43 . The method according to  claim 30 , wherein the second amplification reaction uses a set of primers comprising a primer species that hybridizes to the universal primer site of the adaptor oligonucleotide. 
     
     
         44 . The method according to  claim 30 , wherein the nucleic acid and the adaptor oligonucleotide are double stranded. 
     
     
         45 . The method according to  claim 44 , wherein the double stranded adaptor oligonucleotide comprises a complementary region attached to the nucleic acid and a non-complementary region.