METHODS AND SYSTEMS FOR PROCESSING POLYNUCLEOTIDES
The present disclosure provides methods, compositions and systems for processing polynucleotides from or within individual cells or cell populations. The processing of polynucleotides from or within individual cells or cell populations may be performed by partitioning the individual cells or cell populations with processing reagents for accessing cellular contents, and for uniquely identifying the polynucleotides of a given cell or cell population.
1 . A method, comprising:
(a) providing a reaction volume comprising (i) a cell or cell derivative, and (ii) a bead comprising a barcoded oligonucleotide releasably coupled thereto, wherein said barcoded oligonucleotide is a template switching oligonucleotide; and
(b) releasing said barcoded oligonucleotide from said bead to provide said barcoded oligonucleotide in said reaction volume; and
(c) subjecting said reaction volume to an amplification reaction to generate an amplification product, wherein during said amplification reaction, said reaction volume comprises a template polynucleotide from said cell or cell derivative, said barcoded oligonucleotide and a primer having a sequence towards a 3′ end that hybridizes to said template polynucleotide, and wherein said amplification product has sequence complementarity with said template polynucleotide and said barcoded oligonucleotide.