In order to adopt such a scaled-up effort to track the emergence and divergence of SARS-CoV-2 in New York state, we had to identify the best enrichment PCR protocol, and so we did a lot of searching. We looked and found the ARTIC v3 protocol using the seqWell plexWell method and that seemed highly attractive to us because of the scalability. It actually enabled us to implement low, moderate, and even higher throughput as needed. It wouldn’t have been possible to do without adopting the scalable technology like the plexWell platform.
The seqWell technology has improved efficiency by significantly cutting down operation time on the QC and dilution of cDNA, in addition to pooling of the samples at an earlier timepoint in the process. The plexWell Rapid Single Cell kit has made preparation of high-quality single-cell libraries feasible for regular lab, instead of a formidable task using the alternative methods. Our experience has led to the implementation of the plexWell Single Cell method at Core Facility, expanding their capabilities to support analysis at single cell resolution in our institute.
Key benefits of using plexWell are the timesaving library prep and the easy clean-ups compared to other methods. Being able to create a library in two days allows us to amp up the number of samples we are processing.
The integrated auto-normalization in the plexWell kit allows us to treat 96 samples as 1—one Qubit measurement to get the average concentration of the input plate, one tube to process in the library prep, and one library pool to normalize and load onto the iSeq. This is a huge advantage of the plexWell kit, as it eliminates both the need for individual input DNA purification and individual sample normalization.
I could have saved hours of time-sensitive work if I had used seqWell during my time in the NGS lab!
We have samples of many different species, such as damselflies, bobcats, and rockfish. The kit is flexible across most species we are working with and with a low input amount which is important for threatened species.
Sequencing results obtained for sequencing 192 samples of amplified single-cell cDNA with plexWell™ (blue) and For Nextera™ reagents (orange). Input DNA was pre-normalized, whereas plexWell™ library was made from un-normalized amplified cDNA. Read count variation for plexWell™ showed 27% CV versus 71% CV for Nextera™.