Methods of identifying multiple epitopes in cells
The invention provides methods, compositions, kits and devices for the detection of target molecules. In some embodiments, the invention allows for multiplexed target molecule detection.
1. A method for identifying whether a plurality of targets are present in a plurality of cells comprising: binding to the targets in the plurality of cells a plurality of tags, wherein a tag comprises a unique binding agent (UBA) that is specific for one of the targets, and an epitope specific barcode (ESB) comprising a code that represents the target identity; and subsequently adding multiple assayable polymer subunit (APS) oligonucleotides to each of the bound tags in the plurality of cells in an ordered manner during successive rounds of split pool synthesis wherein the APS oligonucleotides in each round anneal adjacently to the APS from a previous round via an annealing region, and covalently linking the adjacently annealed APS oligonucleotides to each other to create unique codes that represent the identities of individual cells in which the tags are bound, and wherein the method does not include a step of isolating each cell in the plurality of cells.
2. The method of claim 1 , wherein the UBA comprises an antibody.
3. The method of claim 1 , wherein the ESB further comprises a common linker.
4. The method of claim 1 , wherein the ESB comprises a nucleic acid.
5. The method of claim 1 , wherein the adjacent APSs are linked by ligation.
6. The method of claim 1 , wherein the adjacent APSs are linked by Click chemistry.
7. The method of claim 1 , wherein the APSs comprise an amplification primer binding region.
8. The method of claim 1 , wherein annealing region is within an annealing primer.
9. The method of claim 8 , wherein the annealing primer comprises a loop region.