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Fig. 9 | Molecular Neurodegeneration

Fig. 9

From: Long-read sequencing across the C9orf72 ‘GGGGCC’ repeat expansion: implications for clinical use and genetic discovery efforts in human disease

Fig. 9

Schematic of PacBio no-amplification (No-Amp) targeted sequencing. We applied the PacBio no-amplification (No-Amp) Targeted Sequencing method to a C9orf72 G4C2 repeat expansion carrier to better characterize the repeat’s nucleotide content. The No-Amp targeted sequencing method begins with typical SMRTbell library preparation after the target region is excised by restriction enzyme digestion. Cas9 digestion follows with a guide RNA specific to sequence adjacent (Cas9 Cutting Site) to the region of interest (green), leaving the SMRTbells blunt ended. In this case, the guide RNA was specific to sequence upstream (5′) of the G4C2 repeat expansion on the anti-sense strand. A new capture adapter (red) is then ligated to the blunt ends and captured using magnetic beads (magbeads). This process enriches the library for reads containing the region of interest to maximize read depth

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Molecular Neurodegeneration

ISSN: 1750-1326