The PacBio Revio, a cutting-edge DNA sequencer, accelerates research and drives innovation across various sectors.

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About

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PacBio Sequencing

The PacBio Revio, a cutting-edge DNA sequencer, accelerates research and drives innovation across various sectors. It was acquired collaboratively by a consortium of eight leading research institutes, spearheaded by the CSIRO’s Applied Genomics Initiative (AGI) and funded by the Science and Industry Endowment Fund (SIEF). The Revio’s operations are expertly handled by the ANU Biomolecular Resource Facility (BRF) alongside the AGI at CSIRO. This collaboration ensures that the Revio’s sophisticated, high-throughput, long-read sequencing technologies are available to our partners efficiently and cost-effectively. The BRF provides top-notch laboratory services, while the AGI brings a wealth of genomic expertise, guaranteeing optimal outcomes for all users. For inquiries about collaborative opportunities that extend beyond just data generation with the BRF, please reach out to the AGI at agi@csiro.au.

Revio sequencing at the BRF

The BRF offers comprehensive library preparation and sequencing using the CISRO PacBio Revio. In addition to hosting the Revio, the BRF operates and maintains all necessary accessory equipment for the generation of high-quality PacBio libraries. Flagship equipment includes:

-Femto Pulse and Bioanalyzer for checking DNA/RNA size profiles and integrity

-Megaruptor3 for DNA shearing

-Blue pippin for DNA size selection

-10x Chromium iX for single-cell sequencing

You can give us your libraries or submit high-quality nucleic acid extractions for us to do the library preparation. We offer WGS HiFi library preparation as a standard service, and can perform MAS-seq, Kinnex and whole genome amplification sequencing as boutique services.

Sample processing workflow is as follows: 

BRF PscBio

Step 1: Discuss your project with BRF staff:

The quality of data and total output of a Revio run depends mostly on the quality of the input material; hence, the most critical step of the PacBio sequencing workflow is obtaining high quality extractions, which can be very challenging for non-model organisms. We highly recommend following PacBio guidelines for extractions and DNA quality control.

We recommend getting in touch with BRF early in your project to discuss your data needs and the sequencing efforts required to achieve them. Information such as species, sample extraction method, genome size, amount of data required per sample etc., would help mapping the best strategy for you.

Step 2: Submit samples online using bookbrf.anu.edu.au and get them delivered to the BRF

Email brf@anu.edu.au with your lab details. Our staff will create a bookbrf account for your lab and include your designated lab users.  With an active bookbrf account, you can fill the sample submission form that matches your experimental needs to create a traceable an electronic record of your request. You can then bring the samples or send them by courier to us, location and delivery details here.

Step 3: BRF QC checks

We do full QC checks on your samples and will report the results and our assessment on their suitability for PacBio sequencing. We may ask for explicit written authorisation from you for the project to proceed.

Step 4: Library preparation and sequencing

We perform all necessary checks during the library preparations and monitor the sequencing closely. We will inform you on the performance of your project and liaise with vendors when troubleshooting is required.

Due to the often-unpredictable nature of long-read DNA sequencing, BRF will reserve the right to charge for assays in full, even in the event of an unsuccessful outcome, provided that the assay was executed correctly.

Step 5: Data distribution

BRF will share the full run directory (per SMRT cell) with the client, which is predicted to be ~50GB for a 90 Gb SMRTcell. We can deliver data via share links.