Skip to main content
Erschienen in: Microsystem Technologies 2/2017

05.04.2015 | Technical Paper

Genomic DNA extraction from whole blood using a digital microfluidic (DMF) platform with magnetic beads

verfasst von: Ping-Yi Hung, Pei-Shing Jiang, Erh-Fang Lee, Shih-Kang Fan, Yen-Wen Lu

Erschienen in: Microsystem Technologies | Ausgabe 2/2017

Einloggen

Aktivieren Sie unsere intelligente Suche, um passende Fachinhalte oder Patente zu finden.

search-config
loading …

Abstract

DNA extraction is one of the most essential steps in nucleic acid testing, which usually requires a series of reagent mixing, centrifugal separations, and manual operations. While microfluidic devices have shown their capability in point-of-care applications, some of them face the challenge in on-chip sample preparation, such as DNA extraction from blood samples, prior to the diagnosis. Although digital microfluidic (DMF) eliminates the need of complicated fluidic actuation components and draws great attentions as a promising Lab-on-a-Chip platform, its capability in DNA extraction has been limitedly shown. Herein, we develop a protocol on a DMF platform for DNA extraction. As the protocol contains blood, magnetic beads, a variety of liquids, and complicated steps, it was implemented in a sequence of droplet operations with our proposed washing/droplet-separation strategy. Our on-chip extraction protocol was conducted with room-temperature incubations and much less washing cycles. The results were confirmed by using SYBR-Green dye staining and gel electrophoresis. Therefore, the DMF device was shown to permit on-chip DNA extraction with fewer sample/reagent consumption and shorter operation time.

Sie haben noch keine Lizenz? Dann Informieren Sie sich jetzt über unsere Produkte:

Springer Professional "Wirtschaft+Technik"

Online-Abonnement

Mit Springer Professional "Wirtschaft+Technik" erhalten Sie Zugriff auf:

  • über 102.000 Bücher
  • über 537 Zeitschriften

aus folgenden Fachgebieten:

  • Automobil + Motoren
  • Bauwesen + Immobilien
  • Business IT + Informatik
  • Elektrotechnik + Elektronik
  • Energie + Nachhaltigkeit
  • Finance + Banking
  • Management + Führung
  • Marketing + Vertrieb
  • Maschinenbau + Werkstoffe
  • Versicherung + Risiko

Jetzt Wissensvorsprung sichern!

Springer Professional "Technik"

Online-Abonnement

Mit Springer Professional "Technik" erhalten Sie Zugriff auf:

  • über 67.000 Bücher
  • über 390 Zeitschriften

aus folgenden Fachgebieten:

  • Automobil + Motoren
  • Bauwesen + Immobilien
  • Business IT + Informatik
  • Elektrotechnik + Elektronik
  • Energie + Nachhaltigkeit
  • Maschinenbau + Werkstoffe




 

Jetzt Wissensvorsprung sichern!

Literatur
Zurück zum Zitat Boles DJ, Benton JL, Siew GJ, Levy MH, Thwa Prasanna K, Sandahl MA, Rouse JL, Perkins LC, Sudarsan AP, Jalili R, Pamula VK, Srinivasan V, Fair RB, P. B. G. ffi. n, Eckhardt AE, Pollack MG (2011) Droplet-based pyrosequencing using digital microfluidics. Anal Chem 83:8439–8447CrossRef Boles DJ, Benton JL, Siew GJ, Levy MH, Thwa Prasanna K, Sandahl MA, Rouse JL, Perkins LC, Sudarsan AP, Jalili R, Pamula VK, Srinivasan V, Fair RB, P. B. G. ffi. n, Eckhardt AE, Pollack MG (2011) Droplet-based pyrosequencing using digital microfluidics. Anal Chem 83:8439–8447CrossRef
Zurück zum Zitat Claassen S, du Toit E, Kaba M, Moodley C, Zar HJ, Nicol MP (2013) A comparison of the efficiency of five different commercial DNA extraction kits for extraction of DNA from faecal samples. J Microbiol Methods 94:103–110CrossRef Claassen S, du Toit E, Kaba M, Moodley C, Zar HJ, Nicol MP (2013) A comparison of the efficiency of five different commercial DNA extraction kits for extraction of DNA from faecal samples. J Microbiol Methods 94:103–110CrossRef
Zurück zum Zitat Claassena S, Toit ED, Kaba M, Moodley C, Zar HJ, Nicol MP (2013) A comparison of the efficiency of five different commercial DNA extraction kits for extraction of DNA from faecal samples. J Microbiol Methods 94:103–110CrossRef Claassena S, Toit ED, Kaba M, Moodley C, Zar HJ, Nicol MP (2013) A comparison of the efficiency of five different commercial DNA extraction kits for extraction of DNA from faecal samples. J Microbiol Methods 94:103–110CrossRef
Zurück zum Zitat Fan S-K, Wang F-M (2014) Multiphase optofluidics on an electro-microfluidic platform powered by electrowetting and dielectrophoresis. Lab Chip 14:2728–2738CrossRef Fan S-K, Wang F-M (2014) Multiphase optofluidics on an electro-microfluidic platform powered by electrowetting and dielectrophoresis. Lab Chip 14:2728–2738CrossRef
Zurück zum Zitat Geng T, Bao N, Sriranganathanw N, Li L, Lu C (2012) Genomic DNA extraction from cells by electroporation on an integrated microfluidic platform. Anal Chem 84:9632–9639CrossRef Geng T, Bao N, Sriranganathanw N, Li L, Lu C (2012) Genomic DNA extraction from cells by electroporation on an integrated microfluidic platform. Anal Chem 84:9632–9639CrossRef
Zurück zum Zitat Hilz H, Wiegers U, Adamietz P (1975) Stimulation of proteinase K action by denaturing agents application to the isolation of nucleic acids and the degradation of masked proteins. Eur J Biochem 56:103–108CrossRef Hilz H, Wiegers U, Adamietz P (1975) Stimulation of proteinase K action by denaturing agents application to the isolation of nucleic acids and the degradation of masked proteins. Eur J Biochem 56:103–108CrossRef
Zurück zum Zitat Hsieh Y-L, Ho T-Y, Chakrabarty K (2014) Biochip synthesis and dynamic error recovery for sample preparation using digital microfluidics. IEEE Trans Comput Aided Des Integr Circuits Syst 33:183–196CrossRef Hsieh Y-L, Ho T-Y, Chakrabarty K (2014) Biochip synthesis and dynamic error recovery for sample preparation using digital microfluidics. IEEE Trans Comput Aided Des Integr Circuits Syst 33:183–196CrossRef
Zurück zum Zitat Hua Z, Rouse JL, Eckhardt AE, Srinivasan V, Pamula VK, Schell WA, Benton JL, Mitchell TG, Pollack MG (2010) Multiplexed real-time polymerase Chain reaction on a digital microfluidic platform. Anal Chem 82:2310–2316CrossRef Hua Z, Rouse JL, Eckhardt AE, Srinivasan V, Pamula VK, Schell WA, Benton JL, Mitchell TG, Pollack MG (2010) Multiplexed real-time polymerase Chain reaction on a digital microfluidic platform. Anal Chem 82:2310–2316CrossRef
Zurück zum Zitat Hung P-Y, Lu H-Y, Fan S-K, Lu Y-W (2014) Mechatronic systems in digital microfluidics. Int J Autom Smart Technol 4:216–221CrossRef Hung P-Y, Lu H-Y, Fan S-K, Lu Y-W (2014) Mechatronic systems in digital microfluidics. Int J Autom Smart Technol 4:216–221CrossRef
Zurück zum Zitat Jain M, Nijhawan A, Tyagi AK, Khurana JP (2006) Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochem Biophys Res Commun 345:646–651CrossRef Jain M, Nijhawan A, Tyagi AK, Khurana JP (2006) Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochem Biophys Res Commun 345:646–651CrossRef
Zurück zum Zitat Jebrail MJ, Wheeler AR (2010) Let’s get digital: digitizing chemical biology with microfluidics. Curr Opin Chem Biol 14:574–581CrossRef Jebrail MJ, Wheeler AR (2010) Let’s get digital: digitizing chemical biology with microfluidics. Curr Opin Chem Biol 14:574–581CrossRef
Zurück zum Zitat Jebrail MJ, Bartsch MS, Patel KD (2012) Digital microfluidics: a versatile tool for applications in chemistry, biology and medicine. Lab Chip 12:2452–2463CrossRef Jebrail MJ, Bartsch MS, Patel KD (2012) Digital microfluidics: a versatile tool for applications in chemistry, biology and medicine. Lab Chip 12:2452–2463CrossRef
Zurück zum Zitat Jebrail MJ, Sinha A, Vellucci S, Renzi RF, Cesar Ambriz C, Gondhalekar JS Schoeniger, Patel Kamlesh D, Branda SS (2014) World-to-digital-microfluidic interface enabling extraction and purification of RNA from human whole blood. Anal Chem 86:3856–3862CrossRef Jebrail MJ, Sinha A, Vellucci S, Renzi RF, Cesar Ambriz C, Gondhalekar JS Schoeniger, Patel Kamlesh D, Branda SS (2014) World-to-digital-microfluidic interface enabling extraction and purification of RNA from human whole blood. Anal Chem 86:3856–3862CrossRef
Zurück zum Zitat Kim J, Johnson M, Hill P, Sonkul RS, Kim J, Gale BK (2012) Automated microfluidic DNA/RNA extraction with both disposable and reusable components. J Micromech Microeng 22:015007CrossRef Kim J, Johnson M, Hill P, Sonkul RS, Kim J, Gale BK (2012) Automated microfluidic DNA/RNA extraction with both disposable and reusable components. J Micromech Microeng 22:015007CrossRef
Zurück zum Zitat Kim H, Jebrail MJ, Sinha A, Bent ZW, Solberg OD, Williams KP, Langevin SA, Renzi RF, Van De Vreugde JL, Meagher RJ, Schoeniger JS, Lane TW, Branda SS, Bartsch MS, Patel KD (2013) A microfluidic DNA library preparation platform for next-generation sequencing. Plos One 8:e68988CrossRef Kim H, Jebrail MJ, Sinha A, Bent ZW, Solberg OD, Williams KP, Langevin SA, Renzi RF, Van De Vreugde JL, Meagher RJ, Schoeniger JS, Lane TW, Branda SS, Bartsch MS, Patel KD (2013) A microfluidic DNA library preparation platform for next-generation sequencing. Plos One 8:e68988CrossRef
Zurück zum Zitat Mousa NA, Jebrail MJ, Yang H, Abdelgawad M, Metalnikov P, Chen J, Wheeler AR, Casper RF (2009) Droplet-scale estrogen assays in breast tissue, Blood, and Serum. Sci Transl Med 1:1ra2CrossRef Mousa NA, Jebrail MJ, Yang H, Abdelgawad M, Metalnikov P, Chen J, Wheeler AR, Casper RF (2009) Droplet-scale estrogen assays in breast tissue, Blood, and Serum. Sci Transl Med 1:1ra2CrossRef
Zurück zum Zitat Rival A, Jary D, Delattre C, Fouillet Y, Castellan G, Bellemin-Comte A, Gidrol X (2014) An EWOD-based microfluidic chip for single-cell isolation, mRNA purification and subsequent multiplex qPCR. Lab Chip 14(19):3739–3749CrossRef Rival A, Jary D, Delattre C, Fouillet Y, Castellan G, Bellemin-Comte A, Gidrol X (2014) An EWOD-based microfluidic chip for single-cell isolation, mRNA purification and subsequent multiplex qPCR. Lab Chip 14(19):3739–3749CrossRef
Zurück zum Zitat Shamsi MH, Choi K, Ng AHC, Wheeler AR (2014) A digital microfluidic electrochemical immunoassay. Lab Chip 14:547–554CrossRef Shamsi MH, Choi K, Ng AHC, Wheeler AR (2014) A digital microfluidic electrochemical immunoassay. Lab Chip 14:547–554CrossRef
Zurück zum Zitat Shen H-H, Fan S-K, Kim C-J, Yao D-J (2014) EWOD microfluidic systems for biomedical applications. Microfluid Nanofluid 16:965–987CrossRef Shen H-H, Fan S-K, Kim C-J, Yao D-J (2014) EWOD microfluidic systems for biomedical applications. Microfluid Nanofluid 16:965–987CrossRef
Zurück zum Zitat Sista R, Hua Z, Thwar P, Sudarsan A, Srinivasan V, Eckhardt A, Pollack M, Pamula V (2008) Development of a digital microfluidic platform for point of care testing. Lab Chip 8:2091–2104CrossRef Sista R, Hua Z, Thwar P, Sudarsan A, Srinivasan V, Eckhardt A, Pollack M, Pamula V (2008) Development of a digital microfluidic platform for point of care testing. Lab Chip 8:2091–2104CrossRef
Metadaten
Titel
Genomic DNA extraction from whole blood using a digital microfluidic (DMF) platform with magnetic beads
verfasst von
Ping-Yi Hung
Pei-Shing Jiang
Erh-Fang Lee
Shih-Kang Fan
Yen-Wen Lu
Publikationsdatum
05.04.2015
Verlag
Springer Berlin Heidelberg
Erschienen in
Microsystem Technologies / Ausgabe 2/2017
Print ISSN: 0946-7076
Elektronische ISSN: 1432-1858
DOI
https://doi.org/10.1007/s00542-015-2512-9

Weitere Artikel der Ausgabe 2/2017

Microsystem Technologies 2/2017 Zur Ausgabe

Neuer Inhalt