Figure 1.

Overview of single-molecule real-time (SMRT) sequencing technology. (A) Sequencing starts with library preparation of ultra-long double-stranded DNA. In the next step, adapters, DNA polymerase, and primers bind to the double-stranded DNA, creating the SMRT-bell, which will be loaded later to the SMRT-cell. (B) The library is randomly distributed in the SMRT-cell in the sequencer instrument, in the ideal condition one-third of the ZMWs will be loaded by an SMRT-bell. In each ZMW, the DNA polymerase together with an SMRT-bell are bound to the bottom of the ZMW. The SMRT sequencing uses the circular DNA template to generate a continuous long read in each ZMW chamber. Afterwards, the adapters are trimmed from this long read and overlapping reads can be combined to one consensus sequence of high quality called HiFi read.