The Pacific Biosciences (PacBio) Sequel II is the latest generation of long read sequencer from PacBio. We offer a variety of services to take full advantage of PacBio’s proven long-read technology, Single Molecule, Real-Time (SMRT) Sequencing. Contact Katie Carter or Brent Kronmiller to discuss your project and find out how the Sequel II’s technology can work for you.
PacBio offers two general types of sequencing, circular consensus sequencing (CCS) and continuous long read (CLR) sequencing. As described below in PacBio’s glossary of terms, CCS reads results from the consensus of multiple passes of the polymerase over the same DNA molecule. The resulting consensus sequence is highly accurate since errors in individual subreads are not reflected in the consensus sequence. DNA is sheared for most CCS libraries to create libraries with ~10-20 kb fragment sizes that are short enough for the polymerase to pass over multiple times.
CLR sequencing, on the other hand, is optimized to create the longest reads possible. Since multiple passes over the same DNA molecule are not made due to the length of the template, each read reflects one pass of the polymerase over the DNA molecule. Depending on the size of the genomic DNA, shearing may not be necessary for this type of sequencing. Error rates are higher for this method.
CCS reads for whole genome sequencing projects are called HiFi reads. Due to the narrow size selection required to target DNA fragment sizes suitable for CCS sequencing, a higher DNA input is required for HiFi libraries compared to standard or low input whole genome sequencing protocols.
A variety of library preparation options are available for PacBio sequencing at the CQLS:
Whole genome sequencing
Low input options (with as little as 500 ng on input material) and standard options (5 ug of input material) are available. Low input options cannot be sequenced on more than one SMRT cell
Iso-Seq for full-length RNA transcript sequencing
Multiplexed Microbial Libraries
Multiplex up to 16 microbes in one library with our standard pricing, or up to 48 microbes for an additional custom adapter fee.
Shotgun Metagenomic Libraries
Profile up to 4 communities on one SMRT cell
16s rRNA sequencing and targeted amplicon sequencing options are also available. Contact us for custom pricing.