Abstract
Digital PCR (dPCR) is a molecular biology technique going through a renaissance. With the arrival of new instrumentation dPCR can now be performed as a routine molecular biology assay. This exciting new technique provides quantitative and detection capabilities that by far surpass other methods currently used. This chapter is an overview of some of the applications currently being performed using dPCR as well as the fundamental concepts and techniques this technology is based on.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Sykes PJ, Neoh SH, Brisco MJ et al (1992) Quantitation of targets for PCR by use of limiting dilution. Biotechniques 13:444–449
Vogelstein B, Kinzler KW (1999) Digital PCR. Proc Natl Acad Sci U S A 96:9236–9241
Strain MC, Richman DD (2013) New assays for monitoring residual HIV burden in effectively treated individuals. Curr Opin HIV AIDS 8:106–110
Strain MC, Lada SM, Luong T et al (2013) Highly precise measurement of HIV DNA by droplet digital PCR. PLoS One. doi:10.1371/journal.pone.0055943
Kelly K, Cosman A, Belgrader P et al (2013) Detection of methicillin-resistant staphylococcus aureus by a duplex droplet digital PCR assay. J Clin Microbiol 51:2033–2039
Heredia NJ, Belgrader P, Wang S et al (2012) Droplet Digital™ PCR quantitation of HER2 expression in FFPE breast cancer samples. Methods 59:S20–S23
Dodd DW, Gagnon KT, Corey DR (2013) Digital quantitation of potential therapeutic target RNAs. Nucleic Acid Ther 23:188–194
Boettger LM, Handsaker RE, Zody MC et al (2012) Structural haplotypes and recent evolution of the human 17q21.31 region. Nat Genet 44:881–885
Porensky PN, Mitrpant C, McGovern VL et al (2012) A single administration of morpholino antisense oligomer rescues spinal muscular atrophy in mouse. Hum Mol Genet 21: 1625–1638
Gevensleben H, Garcia-Murillas I, Graeser MK et al (2013) Noninvasive detection of HER2 amplification with plasma DNA digital PCR. Clin Cancer Res 19:3276–3284
Nadauld L, Regan JF, Miotke L et al (2012) Quantitative and sensitive detection of cancer genome amplifications from formalin fixed paraffin embedded tumors with droplet digital PCR. Transl Med (Sunnyvale)). Doi: pii: 1000107
Hindson BJ, Ness KD, Masquelier DA et al (2011) High-throughput droplet digital PCR system for absolute quantitation of DNA copy number. Anal Chem 83:8604–8610
Yeh I, von Deimling A, Bastian BC (2013) Clonal BRAF mutations in melanocytic nevi and initiating role of BRAF in melanocytic neoplasia. J Natl Cancer Inst 105: 917–919
Morisset D, Stebih D, Milavec M et al (2013) Quantitative analysis of food and feed samples with droplet digital PCR. PLoS One. doi:10.1371/journal.pone.0062583
Warren L, Bryder D, Weissman IL et al (2006) Transcription factor profiling in individual hematopoietic progenitors by digital RT-PCR. Proc Natl Acad Sci U S A 103:17807–17812
Ottesen EA, Hong JW, Quake SR et al (2006) Microfluidic digital PCR enables multigene analysis of individual environmental bacteria. Science 314:1464–1467
Fan HC, Stephen R (2007) Detection of aneuploidy with digital polymerase chain reaction. Anal Chem 79:7576–7579
Morrison T, Hurley J, Garcia J et al (2006) Nanoliter high throughput quantitative PCR. Nucleic Acids Res 34:e123
Sundberg SO, Wittwer CT, Gao C et al (2010) Spinning disk platform for microfluidic digital polymerase chain reaction. Anal Chem 82:1546–1550
Huggett JF, Foy CA, Benes V et al (2013) The digital MIQE guidelines: minimum information for publication of quantitative digital PCR experiments. Clin Chem 59:892–902
Acknowledgements
I would like to acknowledge and thank my colleagues at Bio-Rad Laboratories; Adam McCoy, George Karlin-Neumann, Jack Reagan, Svilen Tzonev, and all the members of the Digital Biology Center for their contributions to this overview and for their tireless efforts in the development and advancement of digital PCR.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Bizouarn, F. (2014). Introduction to Digital PCR. In: Biassoni, R., Raso, A. (eds) Quantitative Real-Time PCR. Methods in Molecular Biology, vol 1160. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0733-5_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0733-5_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0732-8
Online ISBN: 978-1-4939-0733-5
eBook Packages: Springer Protocols