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.
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References
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 83:8439–8447
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 94:103–110
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 94:103–110
Fan S-K, Wang F-M (2014) Multiphase optofluidics on an electro-microfluidic platform powered by electrowetting and dielectrophoresis. Lab Chip 14:2728–2738
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 84:9632–9639
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 56:103–108
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 33:183–196
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 82:2310–2316
Hung P-Y, Lu H-Y, Fan S-K, Lu Y-W (2014) Mechatronic systems in digital microfluidics. Int J Autom Smart Technol 4:216–221
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 345:646–651
Jebrail MJ, Wheeler AR (2010) Let’s get digital: digitizing chemical biology with microfluidics. Curr Opin Chem Biol 14:574–581
Jebrail MJ, Bartsch MS, Patel KD (2012) Digital microfluidics: a versatile tool for applications in chemistry, biology and medicine. Lab Chip 12:2452–2463
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 86:3856–3862
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 22:015007
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:e68988
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 1:1ra2
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):3739–3749
Shamsi MH, Choi K, Ng AHC, Wheeler AR (2014) A digital microfluidic electrochemical immunoassay. Lab Chip 14:547–554
Shen H-H, Fan S-K, Kim C-J, Yao D-J (2014) EWOD microfluidic systems for biomedical applications. Microfluid Nanofluid 16:965–987
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 8:2091–2104
Acknowledgments
The authors thank Ms. T. Kirk for manuscript editing. The collaborative project has been supported by Industrial Technology Research Institute (ITRI), Taiwan, and Ministry of Science and Technology, Taiwan (MOST 102-2221-E-002-084-MY3).
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Hung, PY., Jiang, PS., Lee, EF. et al. Genomic DNA extraction from whole blood using a digital microfluidic (DMF) platform with magnetic beads. Microsyst Technol 23, 313–320 (2017). https://doi.org/10.1007/s00542-015-2512-9
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DOI: https://doi.org/10.1007/s00542-015-2512-9