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PacBio SMRT Sequencing

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CD Genomics is providing PacBio Single Molecular Real-Time (SMRT) sequencing to complement our NGS facility. By taking advantage of the long-read and single molecular sequencing capability developed by PacBio, we are proud to offer advanced genome de novo assembly solutions and full-length gene/transcript sequencing strategy to suit your project needs.

PacBio SMRT Sequencing

Owing to highly complex of the genomes with many long repetitive elements, copy number alterations, and structural variations, short-read paired-end technologies are insufficient for discovery. Long-read sequencing delivers reads in excess of several kilobases, which can span complex or repetitive regions with a single continuous read, allowing for the resolution of these large structural features. PacBio utilizes SMRT sequencing technology to rapid sequence long-read fragments with high quality, making it well-suited for unsolved problems in genome, transcriptome, and epigenetics research. It combines long read technology and rapid sequencing to produce the highest-quality de novo assemblies, phase alleles and variants, and significantly improve the detection of large structural variants.

This technology employs a specialized flow cell with many thousands of individual picolitre wells with transparent bottoms — zero-mode waveguides (ZMW), the polymerase is fixed to the bottom of the well and allows the DNA strand to progress through the ZMW, as a result, the system can focus on a single molecular. It is also classified as third-generation sequencing (TGS).

CD Genomics are able to provide PacBio SMRT sequencing for several research purposes:

  1. Microbial genome assembly. By employing PacBio SMRT long-read sequencing technology, we can generate a complete genome of bacteria and fungi without gaps or N-based errors. The complete map of a microbe is helpful for a wide range of areas such as pathogen control, industrial fermentation, and functional study, etc.
  2. Full-length small ribosomal subunits 16S/18S/ITS sequencing. 16S rRNA analysis of the variable region is difficult to identify the microorganism to genus level by conventional methods due to the limitation of sequencing read length, we are capable of performing full-length 16S/18S/ITS sequencing, combined with our novel bioinformatics pipeline, to achieve accurate taxonomic identification at the species level. 16S rRNA is usually sequenced to detect prokaryotic microorganisms (total length around 1,500bp), while 18S rRNA is typically sequenced to detect eukaryotic microbes (total length 1,500-2,000bp) and ITS (about 400-900bp).
  3. Metagenomics studies. The long reading sequencing based shotgun metagenomics can provide a clearer and more detailed picture of a microbial community. It provides possibility sequence to a strain-level of the microbial community by constructing different insert length of libraries.
  4. Full-length transcripts sequencing. By taking advantage of PacBio SMRT sequencing technology, people are able to achieve identification and quantification of full-length isoform analysis without the need of fragmentation and post-sequencing assembly, which is useful to identify new transcripts and new introns. Complete transcript from the 5' end to the 3'-poly A tail can be easily obtained with long sequencing reads of 10-15Kb in average, thus accurately identifying isoforms, alternative splicing sites, fusion gene expression, and allelic expression.
  5. Genome-wide DNA modification analysis. PacBio is able to sequences the entire genome region, thus directly obtaining the DNA modification map of a species. The types of DNA modification for accurate PacBio sequencing include 4-mC, 6-mA, and PT (DNA backbone phosphorothioate modifications).

Features and Benefits:

  • Assemble non-model species
  • 10-15 kb average read length
  • No PCR amplification bias
  • Novel bioinformatics analysis programs and pipelines
  • Well-experienced personnel
PacBio SMRT Sequencing
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