The High Pass Plus Cassette
The High Pass Plus™ gel cassette is the newest addition to the Pippin Family. As the name suggests, it is dedicated to our BluePippin “High-Pass” DNA size selection which has been a go-to method for increasing the read lengths for long-read sequencing.
High Pass size selection removes smaller DNA fragments from a sheared genomic DNA (or sequencing library) while collecting the remaining larger fragments above a tightly controlled size threshold. This way, larger molecules can be presented to the detector or droplet, and better sequencing performance can be achieved.
Long-read sequencing sample prep has been improving overall, in terms of set-up time and workflow. We decided to look at the High Pass as well to see if we could optimize the approach. To this end, we designed an entirely new gel cassette dedicated to High Pass– the High Pass Plus. We’re happy to say that we were able to cut the runtime in half and improve performance and yield.
The High Pass Plus cassette has a stocky separation column, so DNA has a shorter distance to travel. The wider column and increased taper provide higher resolution for a cleaner and more accurate size cut-off. The larger sample wells now allow a maximum load of 10ug (100% up from standard Blue Pippin cassettes). We’ve also increased the size of the elution module and surface area of the filtration membrane, bringing about improved sample recovery and reproducibility.
Here’s what we were able to accomplish, when compared to the current BluePippin standard:
• Half the run time
• Twice the loading capacity
• Better recovery and reproducibility
We offer a >15kb High Pass Plus at this time. Next up will be a >20kb and >30kb.