“Long Read” Sequencing
Based on SMRT technology (single molecule real time technology), RSII and Sequel sequencing enables producing very long readings to solve the coverage of low complexity regions and determine structural variants and haplotypes.
A SMRT library is built from fragments of 10 to 20kb. Once the ends have been repaired, single-stranded adapters are attached to them creating a loop (SMRT bell), anchoring point for DNA polymerase (Figure 1).
A complex comprising insert , adapter and polymerase is loaded, immobilised in each well of the sequencing SMRT cells and illuminated by a Zero Mode Waveguides (ZMW). This system detects fluorescent signals in real time during polymerisation (Figure 2).
Sequencing the circularized insert allows to replicate the insert reads to obtain a quality Consensus Sequence (CCS).
Platforms to contact for support of “long reading”sequencing projects
- GeT-PlaGe
- Institut Pasteur-Biomics
- Institut Curie
- Gentyane