Skip to main content

Multiplex Detection of KRAS Mutations Using Passive Droplet Fusion

Part of the Methods in Molecular Biology book series (MIMB,volume 1547)


We describe a droplet microfluidics method to screen for multiple mutations of a same oncogene in a single experiment using passive droplet fusion. Genomic DNA from H1573 cell-line was screened for the presence of the six common mutations of the KRAS oncogene as well as wild-type sequences with a detection efficiency of 98 %. Furthermore, the mutant allelic fraction of the cell-line was also assessed correctly showing that the technique is quantitative

Key words

  • Droplet microfluidics
  • Digital PCR
  • Multiplexing
  • Passive droplet fusion
  • Biomarkers
  • KRAS
  • Cancer

This is a preview of subscription content, access via your institution.

Buying options

USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-4939-6734-6_11
  • Chapter length: 10 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-4939-6734-6
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   139.99
Price excludes VAT (USA)
Hardcover Book
USD   139.99
Price excludes VAT (USA)
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more


  1. Mazutis L, Baret JC, Griffiths AD (2009) A fast and efficient microfluidic system for highly selective one to one droplet fusion. Lab Chip 9(18):2665–2672

    CrossRef  CAS  Google Scholar 

  2. Pekin D, Skhiri Y, Baret JC, Le Corre D, Mazutis L, Ben Salem C, Millot F, El Harrak A, Hutchison JB, Larson JW, Link DR, Laurent-Puig P, Griffiths AD, Taly V (2011) Quantitative and sensitive detection of rare mutations using droplet-based microfluidics. Lab Chip 11(13):2156–2166

    CrossRef  CAS  Google Scholar 

  3. Baret JC, Miller OJ, Taly V, Ryckelynck M, El-Harrak A, Frenz L, Rick C, Samuels ML, Hutchison JB, Agresti JJ, Link DR, Weitz DA, Griffiths AD (2009) Fluorescence-activated droplet sorting (FADS): efficient microfluidic cell sorting based on enzymatic activity. Lab Chip 9(13):1850–1858

    CrossRef  CAS  Google Scholar 

  4. Mazutis, L, BaretJC, Treacy, P, Skhiri, Y, Araghi, AF, Ryckelynck, M, Taly, V, and Griffiths, A. D. (2009) Multi-step microfluidic droplet processing: kinetic analysis of an in vitro translated enzyme. Lab Chip 9(20), 2902–2908

    Google Scholar 

  5. Lievre A, Bachet JB, Le Corre D, Boige V, Landi B, Emile JF, Cote JF, Tomasic G, Penna C, Ducreux M, Rougier P, Penault-Llorca F, Laurent-Puig P (2006) KRAS mutation status is predictive of response to cetuximab therapy in colorectal cancer. Cancer Res 66(8):3992–3995

    CrossRef  CAS  Google Scholar 

  6. Xia YN, Whitesides GM (2006) Soft lithography. Angew Chem Int Ed 37(5):551–575

    Google Scholar 

  7. Clausell-Tormos J, Lieber D, Baret JC, El-Harrak A, Miller OJ, Frenz L, Blouwolff J, Humphry KJ, Koster S, Duan H, Holtze C, Weitz DA, Griffiths AD, Merten CA (2008) Droplet-based microfluidic platforms for the encapsulation and screening of Mammalian cells and multicellular organisms. Chem Biol 15(5):427–437

    CrossRef  CAS  Google Scholar 

Download references


This work was supported by Région Alsace, the Ministère de l’Enseignement Supérieur et de la Recherche, the Université de Strasbourg, the Université Paris Descartes, the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM), the Institut National du Cancer (INCa, no. 2009–1-RT-03-US-1), and the Association pour la recherche sur le Cancer (ARC, no. SL220100601375).

Author information

Authors and Affiliations


Corresponding author

Correspondence to Valerie Taly PhD .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Pekin, D., Taly, V. (2017). Multiplex Detection of KRAS Mutations Using Passive Droplet Fusion. In: Taly, V., Viovy, JL., Descroix, S. (eds) Microchip Diagnostics. Methods in Molecular Biology, vol 1547. Humana Press, New York, NY.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6732-2

  • Online ISBN: 978-1-4939-6734-6

  • eBook Packages: Springer Protocols