No. PacBio does not accept samples for sequencing. See the list of our global certified service providers (CSPs) by geographic region.
Support FAQs
Email support@pacb.com with the name of your employer or institution and your address and we will connect you to the relevant team for your organization.
This video walks you through HiFi sequencing bioinformatics for uncovering germline variation. The talk covers how HiFi reads can provide a better view of genome completeness, an overview of SMRT Link and the GitHub tools that are available for variant calling, information about the PacBio Whole Genome Sequencing variant pipeline, and finally, we take a look at the PacBio compatible partners ecosystem and some helpful additional resources to get you started.
This educational video walks you through the why, what and how of the PacBio Kinnex kits, based on the MAS-Seq method. Specifically, why do you need long reads for RNA sequencing? What are the Kinnex kits? And finally, how do the Kinnex arrays work?
In this video https://www.pacb.com/videos/extraction-to-revio-system-weve-got-you-covered/, you’ll learn about the extraction of high molecular weight DNA, which is optimal for long-read sequencing, the positive correlation that exists between sample quality and sequencing performance, new whole genome and micro lab prep method kits that will scale with your needs, and finally, our compatibility partners who enable sequencing success, all of which is designed with the goal of achieving customizable end-to-end solutions.
To learn more or to order consumables, visit our Consumables page for sample prep and library prep solutions.
You can find additional information on our training where you can search for and download online webinars, e-modules, and other training documents. You can also find various applications (WGS, targeted, epigenetics, RNA and metagenomics) on our sequencing methods page. If you have any questions about our training materials, contact us.
Here is an overview of HiFi applications
You can find the protocols (procedure & checklists) on the protocols page (filtered view from featured documentation).
24 reaction kit: SMRTbell prep kit 3.0 (102-182-700)
96 reaction kit: HiFi prep kit 96 (103-381-200)
Both kits and protocols use the same enzymatic steps and have qualified automated methods available. Enzyme volumes do differ between protocols/kits, and the HiFi prep kit 96 comes with additional SMRTbell cleanup beads for the automated pipette shearing method and additional fill volume to enable up to 4 automated runs of a minimum of 24 samples.
Yes, the Revio HiFi prep kit 96 (103-382-200) and protocol provides an automated solution to all steps of the workflow going from DNA to a sequence–ready library for HiFi systems. Qualified methods are available on the Hamilton NGS STAR MOA system.
HiFi plex prep kit 96. This kit and protocol offer the most cost efficient and streamlined prep for doing microbial WGS on HiFi systems.
The only supported sample type is human blood or cell line extracted with PanDNA or CBB kits. We have been successful internally and at customer sites using other human tissues and extraction kits. Please see our PureTarget app note for more details. To learn more, visit the PureTarget web page.
When using a sample type other than Nanobind-extracted human blood or cell line we recommend the following:
- First, demonstrate success using supported sample types, starting with an 8-plex and increasing sample quantity thereafter.
- Introduce new sample types or extraction methods in limited numbers, for example, 3 or fewer new sample types in an 8-plex of otherwise controls.
- See table 3 in the app note for more information about samples that are officially supported, have been tested, or are not recommended.
Non-human samples are untested and not supported as the guide RNAs used in the panel are designed for the human genome. With non-human samples, you can follow our suggestions on custom guide design. The region captured in the panel is available in Panel coordinate (XLSX).
The PureTarget 20 gene panel is at 5 µM and includes 20 pairs of gRNAs made up of 40 individual gRNAs at 0.125 µM each.
To add on custom genes to the panel, for simplicity, we recommend the following:
- Prepare your custom gRNAs pair per gene at final 5 µM.
- Spike into the PureTarget gRNA mix to prepare custom gRNA mixes prior to making ‘Reaction Mix 2 (RM2)’ in step 3 of the Procedure & checklist – Generating PureTarget repeat expansion panel libraries, while ensuring the total sum molarity of the final mix is 5 µM; the volumes of gRNAs you mix should be proportional to the number of guide pairs, for example add 2 µL of a 1-plex mix to 40 µL of the 20-plex PureTarget panel to keep the sum final molarity at 5 µM.
- Do not change any of the master mix volumes in the protocol.
Please consult with PacBio field application scientist or global service and support for more specific details on library prep
Custom PureParget panels are not supported by PacBio
Custom PureTarget panels are not officially supported by PacBio. Users must design and order their guide RNAs and optimize their custom panels accordingly. PacBio can offer limited guidance on guide RNA design software and strategies for optimizing custom panels. We recommend first demonstrating success on the PureTarget repeat expansion panel using supported sample types before adding new guide RNAs or testing a custom set of guides.
Adding more validated repeat expansion targets: This “flavor” of customization is relatively low risk. Internally, we have demonstrated success in adding up to 5 pairs of guides for additional repeat expansion targets. The design strategy is the same as the fixed 20 gene panel: design guides ~2 kb upstream and downstream of the repeat expansion region with fragments ranging from 3-5 kb.
Adding non-repeat expansion targets: Internally, we have demonstrated success in adding a pair of guides to target a full-length gene with fragment size of ~5 kb, similar in size to the PureTarget repeat expansion panel. Please consult with your field application bioinformatics support scientist for the recommended variant calling tools for your targets.
All other designs like capturing fragments longer than 5 kb or using a tiled design are untested and we do not have any recommendations on design or optimization.
A software tool we recommend for designing guide RNAs is Custom Alt-R™ CRISPR-Cas9 guide RNA from Integrated DNA Technologies (IDT) available here: https://www.idtdna.com/site/order/oligoentry/index/crisprcustom.
We recommend ordering SygRNAs (Synthetic Single Guide RNAs) from MilliporeSigma or sgRNA from IDT rather than the two-part targeting system composed of a separate tracrRNA and crRNA. This way, you only need to order one gRNA and the preparation is simplified (there is no annealing of the two molecules).
PacBio has developed automation methods for extraction on the Thermo Fisher Kingfisher Apex, Flex, and Duo. PacBio has also developed methods for size selection using SRE, shearing with pipette tips, and bead cleanup on the Hamilton Microlab Prep. You can find all of these methods at the third-party equipment webpage. For methods on any other equipment, please reach out to the support team of the appropriate vendor.