Parallel RNA extraction using magnetic beads and a droplet array

被引:41
作者
Shi, Xu [1 ]
Chen, Chun-Hong [1 ,2 ]
Gao, Weimin [1 ]
Chao, Shih-hui [1 ]
Meldrum, Deirdre R. [1 ]
机构
[1] Arizona State Univ, Biodesign Inst, Ctr Biosignatures Discovery Automat, Tempe, AZ 85287 USA
[2] Natl Cheng Kung Univ, Dept Elect Engn, Tainan 70101, Taiwan
关键词
MESSENGER-RNA; SOLID-PHASE; ON-CHIP; PURIFICATION; PLATFORM; DNA; MANIPULATION; MICROCHIP; CELLS;
D O I
10.1039/c4lc01111b
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Nucleic acid extraction is a necessary step for most genomic/transcriptomic analyses, but it often requires complicated mechanisms to be integrated into a lab-on-a-chip device. Here, we present a simple, effective configuration for rapidly obtaining purified RNA from low concentration cell medium. This Total RNA Extraction Droplet Array (TREDA) utilizes an array of surface-adhering droplets to facilitate the transportation of magnetic purification beads seamlessly through individual buffer solutions without solid structures. The fabrication of TREDA chips is rapid and does not require a microfabrication facility or expertise. The process takes less than 5 minutes. When purifying mRNA from bulk marine diatom samples, its repeatability and extraction efficiency are comparable to conventional tube-based operations. We demonstrate that TREDA can extract the total mRNA of about 10 marine diatom cells, indicating that the sensitivity of TREDA approaches single-digit cell numbers.
引用
收藏
页码:1059 / 1065
页数:7
相关论文
共 36 条
[1]   One-step purification of nucleic acid for gene expression analysis via Immiscible Filtration Assisted by Surface Tension (IFAST) [J].
Berry, Scott M. ;
Alarid, Elaine T. ;
Beebe, David J. .
LAB ON A CHIP, 2011, 11 (10) :1747-1753
[2]   Microfluidics-based extraction of viral RNA from infected mammalian cells for disposable molecular diagnostics [J].
Bhattacharyya, Arpita ;
Klapperich, Catherine A. .
SENSORS AND ACTUATORS B-CHEMICAL, 2008, 129 (02) :693-698
[3]   Development of a Low-Resource RNA Extraction Cassette Based on Surface Tension Valves [J].
Bordelon, Hali ;
Adams, Nicholas M. ;
Klemm, Amy S. ;
Russ, Patricia K. ;
Williams, John V. ;
Talbot, H. Keipp ;
Wright, David W. ;
Haselton, Frederick R. .
ACS APPLIED MATERIALS & INTERFACES, 2011, 3 (06) :2161-2168
[4]   Rapid fabrication of microchannels using microscale plasma activated templating (μPLAT) generated water molds [J].
Chao, Shih-hui ;
Carlson, Robert ;
Meldrum, Deirdre R. .
LAB ON A CHIP, 2007, 7 (05) :641-643
[5]   Dynamic DNA hybridization on a chip using paramagnetic beads [J].
Fan, ZH ;
Mangru, S ;
Granzow, R ;
Heaney, P ;
Ho, W ;
Dong, QP ;
Kumar, R .
ANALYTICAL CHEMISTRY, 1999, 71 (21) :4851-4859
[6]   RT-qPCR based quantitative analysis of gene expression in single bacterial cells [J].
Gao, Weimin ;
Zhang, Weiwen ;
Meldrum, Deirdre R. .
JOURNAL OF MICROBIOLOGICAL METHODS, 2011, 85 (03) :221-227
[7]   Magnetic bead handling on-chip: new opportunities for analytical applications [J].
Gijs, MAM .
MICROFLUIDICS AND NANOFLUIDICS, 2004, 1 (01) :22-40
[8]   Multifunctional Picoliter Droplet Manipulation Platform and Its Application in Single Cell Analysis [J].
Gu, Shu-Qing ;
Zhang, Yun-Xia ;
Zhu, Ying ;
Du, Wen-Bin ;
Yao, Bo ;
Fang, Qun .
ANALYTICAL CHEMISTRY, 2011, 83 (19) :7570-7576
[9]  
Guillard R.R., 1975, Culture of Marine Invertebrate Animals, P29, DOI [10.1007/978-1-4615-8714-93, 10.1007/978-1-4615-8714-9_3]
[10]   STUDIES OF MARINE PLANKTONIC DIATOMS .1. CYCLOTELLA NANA HUSTEDT, AND DETONULA CONFERVACEA (CLEVE) GRAN [J].
GUILLARD, RR ;
RYTHER, JH .
CANADIAN JOURNAL OF MICROBIOLOGY, 1962, 8 (02) :229-&