Two Technology Updates From the DNA Sequencing Core

Long Amplicon Sequencing – The UWBC DNA Sequencing Facility is now offering long amplicon sequencing through the same service workflow as our Plasmid Sequencing service. You may submit samples as if they were plasmids ( In order for us to align the data and call variants, you must provide UWBC with the sequences of the forward and reverse primers and a reference sequence for your target of interest. Submitted samples should consist of a single locus amplicon, one clean band, of a 1kb to 8kb or longer PCR product in 10mM Tris or water. The locus may be heterozygous as phasing mutations across the amplicon is possible. Similar to Sanger Sequencing, the resulting data quality is highly dependent on the uniformity of the input PCR product with smears and multiple bands unlikely to generate high quality data. This service is in early development so our ability to analyze fewer uniform amplifications may improve over time.

Updated Sequencing Platforms – The UWBC DNA Sequencing Facility will be updating both the Illumina and Pacific Biosciences sequencers this year.

The Illumina NovaSeq 6000 will be replaced by the NovaSeq X Plus with full implementation of the 1.5B read, 10B read, and 26B read flowcells occurring this fall per Illumina’s roadmap. The greatest impact on pricing will be seen for larger projects run on the 26B read flowcell which will cut sequencing costs per Gb in half.

The PacBio Sequel II systems will be replaced this fall with the Revio. The Revio utilizes 25M well SMRTcells vs the 8M well SMRTcells on the Sequel II so we expect output per SMRTcell to triple. The Revio also has four sequencing stages vs one on the Sequel II so overall throughput will more than quadruple. The large increase in output will result in a roughly 50% reduction in the cost per Gb of sequence after adjusting for slightly higher per SMRTcell pricing.