Fig. 1

Schematic overview of five scRNA-seq methods

Summary of the Tang method, Smart-seq, and the UMI-based sequencing methods STRT-seq, CEL-seq, Drop-seq. Comparative differences of the processes of these methods are outlined: scRNA-seq, reverse transcription, cDNA amplification, purifying and filtration, and library construction. Tang method is the earliest scRNA-seq technology. Single cells are separated by micromanipulation. The overall sequencing sensitivity and accuracy are relative low. In Smart-seq, RNA is reverse transcribed by Moloney mouse leukemia virus(MMLV). The sequencing range can reach the full-length cDNA. It has higher sensitivity and accuracy. STRT-seq and STRT/C1-seq introduce UMI on the basis of Smart-seq and labele with biotin at the 5 ‘end, which can be recovered by magnetic beads. This sequencing method improves the sensitivity and accuracy, but has a strong 5’ end bias. CEL-seq obtains 3 ‘terminal fragment by IVT. The sequencing sensitivity is high, but there is a strong 3’ end bias and the accuracy is low. Drop-seq uses microfluidic technology to package a single cell into an independent droplet, which greatly increases the capture capacity and library capacity of single cell. It has great advantages in detecting a large number of single cell sequencing samples, but the sequencing sensitivity is low