October 28, 2021  |  

SFAF NGS Tech Panel

The COVID-19 pandemic has brought new focus and resources to pathogen surveillance of all kinds. HiFi sequencing, which combines high accuracy, long read lengths, and single-molecule sequencing, is unique in…


August 19, 2021  |  

Technical Note: Preparing DNA for PacBio HiFi sequencing – Extraction and quality control

Single Molecule, Real-Time (SMRT) Sequencing uses the natural process of DNA replication to sequence long fragments of native DNA in order to produce highly accurate long reads, or HiFi reads. As such, starting with high-quality, high molecular weight (HMW) genomic DNA (gDNA) will result in longer libraries and better performance during sequencing. This technical note is intended to give recommendations, tips and tricks for the extraction of DNA, as well as assessing and preserving the quality and size of your DNA sample to be used for HiFi sequencing.


August 19, 2021  |  Products, procedures + protocols

Application Note: Considerations for using the low and ultra-low DNA input workflows for whole genome sequencing

As the foundation for scientific discoveries in genetic diversity, sequencing data must be accurate and complete. With highly accurate long-read sequencing, or HiFi sequencing, there is no longer a compromise between read length and accuracy. HiFi sequencing enables some of the highest quality de novo genome assemblies available today as well as comprehensive variant detection in human samples. PacBio HiFi libraries constructed using our standard library workflows require at least 3 µg of DNA input per 1 Gb of genome length, or ~10 µg for a human sample. For some samples it is not possible to extract this amount of DNA for sequencing. For samples where between 300 ng and 3 ug of DNA is available, the Low DNA Input Workflow enables users to generate high-quality genome assemblies of small-bodied organisms. For samples where even less DNA is available (as low as 5 ng), the amplification-based Ultra-Low DNA Input Workflow is available.


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