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PacBio Sequencing

Achieving a combination of attributes to which all sequencing technologies aim, Pacific Bioscience’s (PacBio) Single Molecule Real-Time (SMRT) sequencing is capable of producing both high per-base accuracy and long read lengths. Known as HiFi reads, the data is generated through circular consensus sequencing (CCS). It is not uncommon for SMRT sequencing to produce reads in excess of 10kb with quality scores above the historical Q30 benchmark.

The SMRT platform detects nucleotides being incorporated onto native DNA strands inside nano-sized cavities called ZMWs (zero mode waveguide) in real-time and for long durations. PacBio’s proprietary CCS approach circularizes input DNA strands with hairpin adapters at each end allowing for continuous, rolling sequencing of the template DNA. The repeated sequencing gives rise to the signature accuracy of HiFi reads, derived from the consensus sequences at each ZMW.

Fragment length is highly controlled on this platform to optimize the number of rotations through each library and accuracy. However, PacBio CCS will commonly produce reads in the 10kb range and is customizable per project.

High-quality data with long read lengths is ideal for resolving large or complex genomes and use cases extend beyond WGS. The PacBio platform is particularly useful for research into isoform discovery and AAV development. However, the platform does require higher-quality samples and may not be the most cost-effective option for smaller genomes.

Each PacBio project has some level of customization. As such, each order must include its own flowcell alongside the appropriate library preparations for each sample, which are indexed to allow for multiplexing on any flowcell. Please explore SeqCenter’s PacBio service offerings below to determine which prep best suits your research needs.

  • PacBio Icon

    PacBio WGS

    PacBio HiFi reads are ideal for WGS of complex, higher-order organisms and improve the likelihood of fully closed genomes. Library preparations are tailored to maximize read lengths for individual sample types, which helps you get the most out of your sequencing.

  • AAV Icon

    PacBio AAV

    Adeno-Associated Virus (AAV) vectors are an increasingly powerful tool for live-host gene therapies. Genomic payloads are delivered by AAV packaged vectors to host tissues to carry out a specific gene edit attributed to a given disease or ailment. Ensuring the accuracy of this genomic template is paramount in generating safe and accessible therapies. As a proud PacBio-sponsored AAV sequencing facility, we are excited to offer library preparation and sequencing for both ssAAV and scAAV samples. The platform’s CCS-generated HiFi reads are a great way to validate vectors and track new mutations.

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    PacBio IsoSeq

    Novel isoform discoveries are within reach with high-quality, full-length mRNA sequencing. PacBio’s unique capabilities are prime for this growing field of study, allowing for detection of novel splicing arrangements possible with no assembly or inference needed. The HiFi reads also guarantee SNP detection for each form.

Other PacBio Applications

While the above services make up SeqCenter’s standard PacBio offerings, we are happy to discuss how our PacBio services may fit into your research with custom library preparations and sequencing projects. PacBio is a great platform for Mas-seq of 10x chromium libraries and full length 16S. The platform also supports a second sequencing approach (in addition to CCS) known as Continuous Long Read sequencing (formerly CLR) that does not produce HiFi reads but instead focuses on generating unparalleled read lengths.

Additionally, PacBio maintains a list of up-to-date citations of studies utilizing their technology that we are happy to explore with you. If you are interested in using SeqCenter’s PacBio services for your project and do not see an approach that is right for you above, please reach out.

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