Outline of PBS. A. Ligations. F-adaptors with F-barcodes at 5' end and R-adaptors are ligated to the miRNAs consequently. B. Reverse transcription. Reverse transcription is carried out by RT-primer to form DNA stand. C. Library amplification. The libraries were amplified by F-primers and R-primers. The R-barcodes are located in the overhangs at 5' end of R-primers, and the right ePCR primers are devised at the 5' end of R-primers. D. ePCR. The ePCR reactions are operated to transfer the libraries to magnetic beads. E. Sequencing. The libraries are sequenced by sequencing primer A and sequencing primer B. The sequencing primer A is in charge of sequencing miRNAs and the F-barcodes while the sequencing primer B is responsible for sequencing R-barcodes.
Tu et al. BMC Genomics 2012 13:43 doi:10.1186/1471-2164-13-43