SAMPLE MULTIPLEXING
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.