Synergistic use of electroosmotic flow and magnetic forces for nucleic acid extraction

被引:17
作者
Deraney, Rachel N. [1 ]
Schneider, Lindsay [1 ]
Tripathi, Anubhav [1 ]
机构
[1] Brown Univ, Ctr Biomed Engn, Sch Engn, 182 Hope St, Providence, RI 02912 USA
关键词
PCR INHIBITORS; DNA; DIELECTROPHORESIS; MICROFLUIDICS; PURIFICATION; SEPARATION; FREQUENCY; PARTICLES; TIME;
D O I
10.1039/c9an02191d
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Nucleic acid sample preparation is essential for biological sample-based diagnostics. It is crucial that diagnostic tests be both specific and sensitive as to provide the most accurate diagnosis possible. Inefficient sample preparation can hinder the specificity and sensitivity of these tests since carryover contaminants can inhibit downstream processes, such as amplification. Microfluidic devices have been used previously to extract nucleic acids from a biological sample due to lower reagent volumes and ease of use. A novel microfluidic chip has been designed for nucleic acid sample preparation which combines electroosmotic flow and magnetic bead-based extraction to isolate DNA from a plasma sample. A steady electric field was incorporated into the microfluidic chip design, which when combined with a glass clover slip and a voltage differential, creates electroosmotic flow. With the goal of isolating nucleic acids into a clean, inhibitor free solution, the electroosmotic flow is the driving force and separation mechanism purifying the DNA sample captured on magnetic beads in the microfluidic chip system. Carryover volume, or the volume of unwanted sample contaminants that accompany the nucleic acids into the final elution buffer, was minimized to 0.22 +/- 0.03%. In combination with magnetic bead based nucleic acid extraction techniques, a 15% increase in DNA extraction yield is reported for the microfluidic chip with the voltage applied versus without. Although the literature on nucleic acid separation in microfluidic chips is abundant, this is the first to combine microfluidic chip design, magnetic bead-based isolation and electroosmotic flow.
引用
收藏
页码:2412 / 2419
页数:8
相关论文
共 23 条
[1]   Current Nucleic Acid Extraction Methods and Their Implications to Point-of-Care Diagnostics [J].
Ali, Nasir ;
Pontello Rampazzo, Rita de Cassia ;
Tavares Costa, Alexandre Dias ;
Krieger, Marco Aurelio .
BIOMED RESEARCH INTERNATIONAL, 2017, 2017
[2]   Dielectrophoresis of DNA: Time- And frequency-dependent collections on microelectrodes [J].
Department of Electronics and Electrical Engineering, University of Glasgow, Glasgow G12 8LT, United Kingdom ;
不详 ;
不详 .
IEEE Trans. Nanobiosci., 2006, 2 (139-146) :1-8
[3]   Magnetic particles for the separation and purification of nucleic acids [J].
Berensmeier, Sonja .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2006, 73 (03) :495-504
[4]   A detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria [J].
Chakravorty, Soumitesh ;
Helb, Danica ;
Burday, Michele ;
Connell, Nancy ;
Alland, David .
JOURNAL OF MICROBIOLOGICAL METHODS, 2007, 69 (02) :330-339
[5]   On slip velocity boundary conditions for electroosmotic flow near sharp corners [J].
Craven, Thomas J. ;
Rees, Julia M. ;
Zimmerman, William B. .
PHYSICS OF FLUIDS, 2008, 20 (04)
[6]   Isolating Influenza RNA from Clinical Samples Using Microfluidic Oil-Water Interfaces [J].
Cui, Francis R. ;
Wang, Jingjing ;
Opal, Steven M. ;
Tripathi, Anubhav .
PLOS ONE, 2016, 11 (02)
[7]   Vortex- and Centrifugation-Free Extraction of HIV-1 RNA [J].
Deraney, Rachel N. ;
Troiano, Derek ;
Joseph, Richard ;
Sam, Soya S. ;
Caliendo, Angela M. ;
Tripathi, Anubhav .
MOLECULAR DIAGNOSIS & THERAPY, 2019, 23 (03) :419-427
[8]   On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids [J].
Garcia-Schwarz, Giancarlo ;
Rogacs, Anita ;
Bahga, Supreet S. ;
Santiago, Juan G. .
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2012, (61)
[9]   Electrokinetic flow control in microfluidic chips using a field-effect transistor [J].
Horiuchi, Keisuke ;
Dutta, Prashanta .
LAB ON A CHIP, 2006, 6 (06) :714-723
[10]   Electric field-induced concentration and capture of DNA onto microtips [J].
Kalyanasundaram, Dinesh ;
Inoue, Shinnosuke ;
Kim, Jong-Hoon ;
Lee, Hyun-Boo ;
Kawabata, Zenko ;
Yeo, Woon-Hong ;
Cangelosi, Gerard A. ;
Oh, Kieseok ;
Gao, Dayong ;
Lee, Kyong-Hoon ;
Chung, Jae-Hyun .
MICROFLUIDICS AND NANOFLUIDICS, 2012, 13 (02) :217-225