A METHOD FOR SINGLE CELL SEQUENCING OF MIRNAS AND OTHER CELLULAR RNAS

摘要

Provided herein are methods for single cell sequencing of miRNAs and other low abundance and/or or short cellular RNAs. A method is provided for preparing an RNA sample for sequencing comprising: (a) obtaining a sample comprising RNA molecules; (b) self-ligating each RNA molecule in the sample to form circular RNA; (c) hybridizing a first set of random primers to the circular RNA; (d) extending the first set of random primers; (e) self-ligating the cDNA to form a circular cDNA; (f) hybridizing a second set of random primers to the circular cDNA; and (g) extending the second set of random primers hybridized to the circular cDNA to form double-stranded cDNA. In some aspects, steps (c) and (d) and/or steps (f) and (g) may be performed simultaneously. In other aspects, steps (c) and (d) and/or steps (f) and (g) may be performed sequentially in the absence of exogenous manipulation.