Long Amplicon Analysis (LAA)
Based on PacBio Circular Consensus Sequences (CCSs), polymerase could copy the same DNA region for several times, which generates high-fidelity, long reads (>99.9% single-molecule read accuracy). Long Amplicon Analysis (LAA) using Single Molecule, Real-Time (SMRT) Sequencing and the Sequel System produces highly accurate and phased CCSs from long amplicons.
Figure 1. Overview of SMRT Sequencing Technology. (From Simon Ardui, et al., 2018)
Compared to the short-read sequencing platforms, PacBio’s long read sequencing makes it is straightforward to sequence amplicons or captured fragments ranging in size from several hundred base pairs up to 10 Kb. These long sequences are very useful for visualization of variants including SNPs, CNVs, and other structural variants that typically do not require assembly.
SeqInsight provides professional and cost-effective LAA service with sequencing depth ranging from <500 to 10K CCSs per sample to meet your specific research requirements.
- Full-length 16S/18S/ITS gene sequencing
- De novo assembly of targeted gene
- Custom amplicon sequencing
- 2 µg or more of amplicons (i.e. purified PCR Products) with a concentration of ~50ng/µl or higher, be dissolved in Tris-HCl buffer (10 mM, pH 8) and shipped on dry ice packs.
- We also provide amplicon preparation service, from starting materials including genomic DNA, cell pellets, and tissues.
For full-length 16S/18S/ITS gene sequencing:
- OTUs clustering and filtering
- OTUs analysis and species annotation
- Alpha diversity, Beta diversity, Meta-analysis
- Multi-variate statistical analysis
For other applications, please inquire.
- Simon Ardui, et al. (2018) Single molecule real-time (SMRT) sequencing comes of age: applications and utilities for medical diagnostics. Nucleic Acids Research. 46(5) 2159–2168.
- Joshua P. Earl, et al. (2018) Species-level bacterial community profiling of the healthy sinonasal microbiome using Pacific Biosciences sequencing of full- length 16S rRNA genes. Microbiome. 6.
- Henk P.J. Buermans, et al. (2017) Flexible and Scalable Full-Length CYP2D6 Long Amplicon PacBio Sequencing. Human Mutation. 38(3) 310–316.