Absolute Quantification
with Droplet Digital PCR

Counting DNA, molecule by molecule

Bio-Rad QX200

Understand how Droplet Digital PCR (ddPCR) has become embedded in molecular biology laboratories that require accurate nucleic acid quantification. The links below guide you through multiple layers of information on absolute quantification with droplet digital PCR. Each layer illustrates how ddPCR has been used to improve precision and accuracy in nucleic acid quantification, thereby ensuring that molecular biology labs can have confidence in their results.

The Pathway to Accuracy

The Pathway to Accuracy

Metrology laboratories across the globe have adopted and validated digital PCR for accurate and precise measurement of nucleic acids. Manufacturers of nucleic acid standards and reference materials have embraced ddPCR for accurate quantification of nucleic acids and variant allele frequencies. External quality assessment (EQA) and proficiency testing (PT) organizations recognize that ddPCR provides a level of robustness for nucleic acid quantification in the clinical environment. It is accepted as the gold standard for validating alternative nucleic acid quantification methods. Ultimately, clinical laboratories are adopting ddPCR for its precision, accuracy and robust workflow.

Clinical Laboratories

Droplet Digital PCR delivers peace of mind in your nucleic acid quantification results. Absolute quantification of nucleic acids and industry-leading precision in variant allele frequency and copy number determination are standard with ddPCR.

Visit the Bio‑Rad publication database to review examples of how ddPCR is improving clinical and translational research and testing.

Example publication from the Bio-Rad ddPCR publication database

Title TERT, BRAF, and NRAS mutational heterogeneity between paired primary and metastatic melanoma tumors
Date 9 Feb, 2020
Journal J Invest Dermatol
Author Chang, GA et al.

Search the Bio‑Rad ddPCR publication database

  1. Go to https://www.bio-rad.com/ddPCR/publications
  2. Under Application Type, select Liquid Biopsy or Cancer from the dropdown menu

Example publication from the Bio-Rad ddPCR publication database

Title Comparison of real-time quantitative PCR and Digital Droplet PCR for BCR-ABL1 monitoring in patients with chronic myeloid leukemia
Date 24 Oct, 2019
Journal J Mol Diagn
Author Franke, GN et al.

Search the Bio‑Rad ddPCR publication database

  1. Go to https://www.bio-rad.com/ddPCR/publications
  2. Under Keyword, type in BCR-ABL

Example publication from the Bio-Rad ddPCR publication database

Title Plasma AR status and cabazitaxel in heavily treated metastatic castration-resistant prostate cancer
Date 1 Jul, 2019
Journal Eur J Cancer
Author Conteduca, V et al.

Search the Bio‑Rad ddPCR publication database

  1. Go to https://www.bio-rad.com/ddPCR/publications
  2. Under Application Type, select Copy Number Variation from the dropdown menu
  3. Under Keyword, type in Cancer

Example publication from the Bio-Rad ddPCR publication database

Title Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets
Date 20 May, 2020
Journal Emerg Microbes Infect
Author Liu, X et al.

Search the Bio‑Rad ddPCR publication database

  1. Go to https://www.bio-rad.com/ddPCR/publications
  2. Under Keyword, type in COVID

Example publication from the Bio-Rad ddPCR publication database

Title Ultra-deep next-generation sequencing of plasma cell–free DNA in patients with advanced lung cancers: results from the Actionable Genome Consortium
Date 20 Mar, 2019
Journal Ann Oncol
Author Li, BT et al.

Search the Bio‑Rad ddPCR publication database

  1. Go to https://www.bio-rad.com/ddPCR/publications
  2. Under Application Type, select Next Generation Sequencing from the dropdown menu

Standards Companies

Reference Materials and Controls
When your business is standards and reference materials, the technology of choice for nucleic acid quantification is Droplet Digital PCR by Bio-Rad.

Droplet Digital PCR delivers peace of mind in nucleic acid quantification. If your controls and reference materials are accurately quantified by ddPCR, then you can rest assured that your clinical and research measurement values are accurate and precise.

Proficiency Testing and Quality Assurance

Proficiency Testing (PT) and External Quality Assessment (EQA)
All clinical labs must participate in external quality assessment programs or proficiency tests for each of their provided clinical tests. The use of Droplet Digital PCR can be beneficial at several stages for providers of PTs and EQAs.

First, in the generation of control materials for distribution across interlaboratory groups, accurate and precise quantification of nucleic acids is essential.

Title One byte at a time: evidencing the quality of clinical service next generation sequencing of germ line and somatic variants
Date March 2020
Journal Eur J Human Gen
Author Gutowska-Ding, M et al.

Second, for EQAs/PTs that generate nucleic acid material as an output (for example, nucleic acid extraction), quantification of those materials by ddPCR provides unequivocal results for interlab comparison.

Interested in learning more? Speak with our sales specialist!

Metrology Laboratories

Metrology: The Science of Measurement
Metrology labs globally have adopted Droplet Digital PCR with a focus on improving the precision and accuracy in measuring nucleic acids.

All national institutes of metrology have a common metrological focus to improve measurement processes and practices in science. However individual research areas differ across the globe and between NMIs.

Application areas that have benefited from metrological support with ddPCR are:

Please hover over to view the reference.

GMO

Virology

Oncology

  1. Kosir AB et al. (2019). Digital PCR as an effective tool for GMO quantification in complex matrices. Food Chem 294, 73–78.
  2. Grohmann L et al. (2017). Screening for six genetically modified soybean lines by an event-specific multiplex PCR method: Collaborative trial validation of a novel approach for GMO detection. J Consum Prot Food Saf 12, 23–36.
  1. Whale AS et al. (2016). Detection of rare drug resistance mutations by digital PCR in a human influenza A virus model system and clinical samples. J Clin Microbiol 54, 392–400.
  2. Cleveland MH et al. (2018). Certification of Standard Reference Material 2365 BK Virus DNA Quantitative Standard. National Institute of Standards and Technology. U.S. Department of Commerce. NIST Standard Reference Materials Publication 260-191.
  3. Dobnik D et al. (2019). Accurate quantification and characterization of adeno-associated viral vectors. Front Microbiol 10, 1,570.
  1. He H-J et al. (2019). Development and interlaboratory evaluation of a NIST reference material RM 8366 for EGFR and MET gene copy number measurements. Clin Chem Lab Med 57, 1,142–1,152.
  2. Whale AS et al. (2018). Assessment of digital PCR as a primary reference measurement procedure to support advances in precision medicine. Clin Chem 64, 1,296–1,307