Everything You Need to Know About Droplet Digital PCR

Droplet Digital polymerase chain reaction (ddPCR™) was developed to provide high-precision, absolute quantification of nucleic acid target sequences with wide-ranging applications for both research and clinical diagnostic applications. ddPCR measures absolute quantities by counting nucleic acid molecules encapsulated in discrete, volumetrically defined water-in-oil droplet partitions. Droplet Digital PCR using Bio-Rad’s QX100™ or QX200™ Droplet Digital PCR system overcomes the previous lack of scalable and practical technologies for digital PCR implementation.

ddPCR has the following benefits for nucleic acid quantification:

• Unparalleled precision — the massive sample partitioning afforded by ddPCR enables small fold differences in target DNA sequence between samples to be reliably measured
• Increased signal-to-noise — enrich for rare targets by reducing competition that comes from high-copy templates
• Removal of PCR efficiency bias — error rates are reduced by removing the amplification efficiency reliance of PCR, enabling accurate quantification of targets
• Simplified quantification — a standard curve is not required for absolute quantification

Download the Droplet Digital™ PCR Applications Guide for a comprehensive explanation of this powerful technique.

The guide includes chapters on:

1. Designing Droplet Digital™ PCR Experiments
2. Absolute Quantification and the Statistics of Droplet Digital™ PCR
3. Copy Number Variation Analysis
4. Rare Mutation and Sequence Detection
5. Gene Expression
6. Next-Generation Sequencing Library Analysis
7. Droplet Digital™ PCR Tips, Assay Considerations, and Troubleshooting

Automated Droplet Generator Saves Time and Improves Data Quality

Bio-Rad Laboratories announced the launch of its Automated Droplet Generator (AutoDG™ Instrument) for digital PCR. Combined with the QX200™ Droplet Reader and a laptop computer with QuantaSoft™ Software, the QX200™ AutoDG™ Droplet Digital™ PCR System provides a worry-free, automated way to generate droplets for high-quality data.

“With a quick setup, you’re able to walk away and return to a full plate of droplets ready for thermal cycling and analysis,” said Carolyn Reifsnyder, Bio-Rad marketing manager, Digital Biology Center.

Generating droplets with a hand pipet can be labor intensive and time consuming, and variations in pipetting habits among different users can cause some degree of inconsistency in the resulting data. Bio-Rad’s AutoDG Instrument creates droplets quickly, reproducibly, and reliably. At maximum capacity, the system can generate droplets for 96 wells in less than 45 minutes.

“Droplet Digital PCR is sensitive enough to detect differences in people’s pipetting habits,” said Viresh Patel, Bio-Rad senior marketing manager, Digital Biology Center. “This instrument eliminates that variability.”

The AutoDG Instrument has its own hood and HEPA filter, reducing contamination so researchers can create droplets on a standard laboratory bench, without the need for a PCR cabinet or cleanroom. Adding to the instrument’s ease of use is a color touch-screen interface and deck lighting that guides the operator through system setup to ensure no steps are forgotten. Confirmation is provided once the required consumables are loaded correctly and 45 minutes later the droplets are ready.

Bio-Rad’s AutoDG Instrument can generate droplets for fluorescent probe–based detection or EvaGreen dye–based detection. The instrument is compatible with the QX100™ and QX200 Droplet Digital PCR Systems.

For more information on Bio-Rad’s AutoDG Droplet Digital PCR System, please visit www.bio-rad.com/AutoDGpr.

Droplet Digital™ PCR Tips & Tricks: ddPCR™ Assay Design

Introduction to Droplet Digital™ PCR: Workflow and Applications

Droplet Digital™ PCR System Wins Scientists’ Choice Award for Best New Life Sciences Product of 2013

Bio-Rad Laboratories, Inc.’s QX200 Droplet Digital PCR (ddPCR™) System was awarded the Scientists’ Choice Award for Best New Life Sciences Product of 2013. The award, presented by SelectScience, recognizes the technology that has most significantly impacted life science research in 2013.

“We are seeing more researchers embrace Droplet Digital PCR for a multitude of applications and to have the QX200 System recognized by SelectScience’s scientific community is a great reflection of that,” said Viresh Patel, senior marketing manager with Bio-Rad’s Digital Biology Center.

More than 250,000 members of the SelectScience community were invited to nominate and vote for their favorite new products of 2013. The award was announced at the American Association for Cancer Research (AACR) annual meeting held April 5–9 in San Diego, CA (watch the video). This was a fitting event for the announcement, as the number of cancer research papers that cite ddPCR nearly quintupled from 2012 to 2013.

Moreover, one of the more talked about presentations at AACR was given by ddPCR users, Dr. Pasi Jänne and Dr. Cloud Paweletz of the Belfer Institute for Applied Cancer Science at the Dana-Farber Cancer Institute.

Also at the meeting, Bio-Rad highlighted promising cancer research applications of ddPCR and previewed the new Automated Droplet Generator and QX200 AutoDG™ Droplet Digital PCR System (watch the video). The Automated Droplet Generator simplifies the ddPCR workflow, enabling generation of droplets for 96 samples in less than 30 minutes. Using the AutoDG System, researchers can minimize hands-on time and eliminate user-to-user variability.

This technology has the potential to benefit labs considerably. “With the arrival of the AutoDG System, we hope to make digital PCR both scalable and practical, especially for labs looking to increase the throughput of their ddPCR-based research and translate their discoveries to the clinic.” said Patel.

To learn more about ddPCR, please visit www.bio-rad.com/QX200.