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 nucleic acid targets is present in a plurality of cells comprising:
a) binding to the nucleic acid targets in the plurality of cells a plurality of unique binding agent (UBA) nucleic acid tags;
b) extending the UBAs bound to the targets, and
c) assembling cell originating barcodes (COB) on the extended UBAs by subsequently adding multiple assayable polymer subunit (APS) oligonudeotides to each of the extended UBAs in the plurality of cells in an ordered manner during successive rounds of split pool synthesis wherein the APS oligonudeotides in each round anneal to the APS from a previous round and are covalently linked to the adjacently annealed APS 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 unique code comprises UBA and COB wherein COB consists of multiple APSs.
3. The method of claim 1 , wherein the code is read by sequencing.
4. The method of claim 1 , wherein the UBA is an antisense DNA for a target mRNA.
5. The method of claim 1 , wherein the APS oligonudeotides in each round anneal to the APS oligonudeotide from the prior round via a shared region of complementarity.
6. The method of claim 1 , wherein the APS oligonudeotides in each round are linked to the APS oligonudeotide from the prior by ligation.