R2C2 protocol using 10x Single-Cell RNA-seq AssaycDNA v1

This protocol is based on the published Rolling Circle to Concatemeric Consensus (R2C2) method (1). The aim is to make elongated cDNA via rolling circle amplification using cDNA generated from the 10x Chromium 3' (or 5') single-cell RNA-seq assay to increase the accuracy of downstream sequencing on an Oxford Nanopore Technologies sequencing platform. The major modification from the original protocol is to add an additional cDNA re-amplification step (see section "10x cDNA re-amplification") to enrich for fragments with the proper sequence (poly(T) at 3' and TSO at 5') at both ends, which is essential in circularization with the 10x Splint fragment.