Integrated Genomics

PacBio Sequel Ile System

Sequel IIeWe are equipped with the Sequel IIe System (PacBio). The Sequel IIe system provides an access to high throughput, cost effective, highly accurate long-read sequencing. The system uses the Single Molecule, Real-Time (SMRT®) Sequencing, a proprietary technology proven to produce highly accurate long reads, known as HiFi reads. Because HiFi reads are extremely accurate, downstream analysis is simplified and streamlined, requiring less compute time than the error-prone long reads of other technologies.

The Benefits of HiFi Reads:

  • Long read lengths up to 25 kb
  • High read accuracy >99%
  • Easy library preparation
  • Lower coverage requirements
  • Small file sizes to minimize compute time
  • A single technology solution for a range of applications

We offer complete PacBio library prep, Blue Pippin size selection and sequencing services. The total output in Gb per SMRT cell depends on the library insert size and DNA quality. For long insert libraries, we see high yield, 32-35 Gb per 8M SMRT cell, but only in case of high molecular weight, high quality DNA.

Although the BluePippin-based size selection is optional, it helps to minimize the number of DNA fragments shorter than 10 kb. So, we highly recommend it for some applications, such as whole genome sequencing and de novo assembly.

Sequel Ile Output Files

The output files from the Sequel IIe system do not include low-quality reads. We will provide you with the following files:

  • .hifi_reads.fastq.gz - Gzipped FASTQ file containing HiFi reads
  • .hifi_reads.fasta.gz - Gzipped FASTA file containing HiFi reads
  • .hifi_reads.bam - BAM file containing HiFi reads
  • .hifi.reads.bam.pbi - Index file that allows for random access of HiFi reads in the BAM file

If 5mC CpG Detection is performed, the following additional files are output:

  • .5mc_report.json
  • .primrose.log

For more information, visit the PacBio Sequel System website.

If you have questions about DNA extraction for PacBio sequencing, please contact ExtractDNA@pacb.com

For general considerations, please check out:

DNA Prep Tech Note (pdf)

Sample Prep Tech Note (pdf)