PacBio Whole Genome Sequencing
SeqCenter PacBio HiFi services can accommodate organisms of any size on the Sequel IIe platform. We recommend PacBio services for large-scale, eukaryotic genomes (amphibians, birds, reptiles, mammals, etc.) at a target depth of coverage between 15-20x.
Using Megaruptor 3 technology and PacBio’s SMRTbell v3.0 library preparation kit, sample gDNA is fragmented to a targeted 15-18kb range and sequenced using the latest PacBio SMRTbell chemistry.
Flowcells and library preparations are ordered separately. To mutliplex samples, please order additional library preparations for each sample to be included.
Our HMW DNA Extraction add-on service is highly recommended as PacBio sequencing is highly sensitive to sample degradation and freshly extracted DNA produces the best results. If this option is not selected, please ship frozen samples on dry ice.
PacBio Sample Requirements:
- Total volume of 60µL or more
- Concentration greater than 35ng/uL as measured by Qubit or other fluorimetric method (not Nanodrop)
- Average genomic fragment size must be greater than 15kb
- Eluted into water or dilute Tris/TE buffer
What You Will Receive:
- Raw Fastq file for each sample
- Bam file, containing run metric data
- Materials and Methods Summary
- Data shared via Box folder within 2 weeks.
Full 8M Flowcell x 1
Run Across a Full Flowcell. Library preparation separate.
Full 8M Flowcell x 2
Run Across 2 Full Flowcells. Library preparation separate.
Full 8M Flowcell x 3
Run Across 3 Full Flowcells. Library preparation separate.
PacBio WGS Library Preparation
Includes QC and barcoding. One needed per sample. Sequencing separate.
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Additional Options for PacBio: HMW DNA Extractions
HMW Genomic DNA isolation is available at a rate of $20 per sample
- Extractions will be performed using Zymo Quick-DNA HMW MagBead Kits
- We do not accept live BSL3 or above samples for extractions.
- Samples should be sent as sealed pre-grown agar plates or aspirated frozen cell pellets (10^9 cells recommended).
Custom PacBio Sequencing Projects
PacBio sequencing technology unlocks a large number of applications aside from whole genome assemblies and we are able to take on projects that are tailored to your needs. Our most popular projects include full length AAV Sequencing, MAS-Seq of 10x Chromium libraries, and CCS services.
Assembly and annotation* services are available for some sample types, including collapsed-genome assembly services(via Flye), or partially- or fully-phased** assembly services (via Hifiasm.)
Please contact us with additional project details for pricing and availability.
*Annotation is conditional to database availability and requires a genus/species NCBI taxonomic ID. Assembly and annotation are required for methylation calls.
**Fully-phased assemblies via Hifiasm (trio-binning) require parental Illumina short reads. Externally generated short read data may be subject to fees if reformatting is required for analysis.