PacBio AAV

We are proud to be a PacBio-sponsored adeno-associated virus (AAV) sequencing facility and to take part in this growing field where accuracy is key to the development of new therapeutics. PacBio’s SMRT platform with CCS is ideal for identifying mutations, truncations, and host integration events in AAV genome populations. We use PacBio’s protocol for preparing multiplexed AAV SMRTbell libraries using the SMRTbell prep kit 3.0, which accommodates both self-complementing (scAAV) and single-stranded (ssAAV) constructs within a sample. Once in double-stranded form, barcoded adaptors are added onto all fragments for multiplexed sequencing.

Because AAV nucleic acids may be in multiple forms in a sample and dsDNA is required for successful sequencing, it is critical that the sample form is disclosed. This allows us to address any non-complementary and self-complementary regions with interior inserted repeats before final library preparation.

Sample Requirements:

  • Minimum of 500 ng total material
  • Maximum volume of 50 µL
  • Must declare if sample is ssAAV, scAAV, or both
  • No contact with Axygen consumables, which interfere with SMRTbell prep
  • Must be submitted on dry ice

What You Will Receive:

  • One fastq file for each sample
  • One bam file with run metric data
  • Sequencing stats summary
  • Materials and methods summary
  • Data shared via Box folder within 2 weeks

Packages & Pricing

Sequencing Package Price*
Revio 25M Flowcell
Run across a Full Flowcell. Library preparation separate.
PacBio AAV Library Preparation
Includes QC and barcoding. One needed per sample. Sequencing separate.
*Orders >48 Samples receive 5% discount

SeqCenter offers convenient AAV extractions using ThermoFisher Scientific’s PureLink Viral RNA/DNA Mini kit. For SeqCenter to perform this service, a minimum of 7×1011 vector must be submitted to meet the 500 ng requirement. Shipping must include dry ice to preserve the sample. For more information and pricing for our AAV extraction service, please contact us.

We offer AAV analysis using the open-source tool Magdoll, which is in active development by PacBio affiliated creators. A note of caution, this tool is not directly supported by PacBio itself and has not yet been peer reviewed.

In order to perform this type of analysis, the AAV backbone sequence (as a fasta file), the coordinate position from ITR to ITR, and host genome NCBI accession number must be provided. Optionally, providing the helper and Rep-Cap plasmid sequences (as fasta files) and coordinate location of the trans plasmid from ‘Rep’ to ‘Cap’ will add more information to the analysis output. The final report is delivered in pdf format.

For additional information and pricing, please contact us.

Go to Top