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.
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.