Droplet Generation Oil

In droplet digital PCR, droplet generation oil plays a crucial role in the process of partitioning the PCR reaction mixture into thousands or millions of individual droplets. These droplets serve as discrete reaction vessels, each containing a fraction of the target DNA or RNA molecules.

Droplet generation oil is a specialized oil-based fluid that is used to create and stabilize the droplets in dPCR. The oil is immiscible with the PCR reaction mixture, forming a distinct phase that surrounds and encapsulates the reaction mixture droplets. This oil barrier helps to isolate and compartmentalize each PCR reaction, preventing cross-contamination and ensuring accurate quantification of target nucleic acids.

The key characteristics of RainSure droplet generation oil include:

1. Immiscibility: The droplet generation oil is immiscible with the PCR reaction mixture. This ensures that the oil forms distinct droplets, separating them from the reaction mixture droplets.

2. Stability: The oil is stable and maintain its physical properties during the PCR cycling process. It does not evaporate, degrade, or cause interference with the PCR reaction.

3. Optical Transparency: The oil has high optical transparency to enable efficient detection and analysis of the fluorescent signals generated during dPCR.

4. Inertness: The oil is chemically inert to avoid interfering with the PCR reaction and DNA amplification.

During the dPCR process, the droplet generation oil and reaction mixture are emulsified. This emulsification process creates a stable suspension of droplets, with each droplet containing a single target molecule or none at all. The droplets generated with the help of the droplet generation oil are subsequently subjected to thermal cycling, where the PCR amplification takes place. After amplification, the droplets are individually analyzed to determine the presence or absence of the target molecule based on fluorescence signals.

In summary, droplet generation oil is a critical component in droplet digital PCR as it enables the formation and stabilization of individual droplets containing the PCR reaction mixture. It allows for the precise partitioning and isolation of target nucleic acids, facilitating accurate quantification and analysis in dPCR assays.