QPCR Core

Our qPCR equipment consists of 3 QuantStudio3 qPCR machines and an Opticon2. Software for qPCR setup and analysis is freely available, and includes many routinely used protocols:

• Standard Curve
• Relative Standard Curve
• ΔΔCt
• Melt Curve
• Genotyping
• Presence/Absence

The QuantStudio3 machines have 4 channels for excitation and emission covering many commonly used dye sets, including SYBR green and TaqMan probes. They have also been used for compatible protocols such as Thermal Shift Assay using SYPRO Orange.

For more precise applications, the core houses a QX200 AutoDG ddPCR suite. Droplet digital PCR (ddPCR) was developed to overcome Real-time PCR limitations. It partitions a sample into thousands to tens of thousands of parallel PCR reactions which are categorized by fluorescent signals as positive or negative, providing a digital readout. This method enables absolute quantification without a standard curve. The QX200 system partitions 20uL samples into 20,000 reactions, offering wide dynamic range, precision, and sensitivity while minimizing optimization and validation needs.

Droplet Digital PCR offers unparalleled accuracy, precision, and sensitivity in quantifying DNA molecules. Its applications span copy number variation, mutation detection, absolute quantitation, rare sequence detection, gene expression analysis, and beyond.

Compared to conventional real-time PCR systems, Droplet Digital PCR offers:

• Unparalleled precision, 10x greater precision than real-time PCR, (+/- 10%)
• Detection of rare targets in complex backgrounds, 100x more sensitive than real-time PCR
• Absolute and Direct concentration measurement reported in Copies/ul
• Standard curves are not required with Droplet Digital PCR and no more Ct or ΔΔCT values.