Abstract

Tube-strip digital PCR: fast, easy and Clarity

Johnson Ng, Founder And Chief Executive Officer, JN Medsys Pte Ltd

Sample partitioning, an important step in digital PCR, is typically carried out either using specialised chips, or through generating droplets in an oil emulsion. Chip-based partitions are more stable and since partitions are imaged concurrently, detection times are also shorter. However, the multiple steps required for handling the chips often result in a more cumbersome workflow. Chips might also require dedicated thermal cyclers which might limit the number of reactions that can be performed in a single experiment. Droplets in an oil emulsion, on the other hand, can be carried out using conventional PCR well-plates. These usually have an easier workflow and more reactions can be carried out concurrently. Droplets, though, can be more unstable, and detection times also tend to be longer. Here, we introduce the ClarityTM digital PCR system which combines the best of both chip- and droplet-based partitioning. Sample partitioning is performed on a high-density chip within a conventional PCR tube-strip format. This provides stable chip-based partitions yet allows users to handle only the familiar PCR tube-strips. Sample partitioning requires just 1-2 min and up to 96 reactions can be carried out in a single experiment. The tube-strips are also compatible with many existing thermal cyclers. Workflow is fast and easy, with the flexibility to increase the reaction throughput. On the ClarityTM, existing qPCR assays are also easily adapted into digital PCR with minimal optimisation. Our aim is to make performing digital PCR as easy as qPCR, and the ClarityTM is one step in that direction.