Magnetic-bead-based microfluidic system for ribonucleic acid extraction and reverse transcription processes

被引:42
作者
Liu, Chien-Ju [1 ]
Lien, Kang-Yi [2 ]
Weng, Ching-Yi [3 ]
Shin, Jyh-Wei [4 ]
Chang, Tsuey-Yu [4 ]
Lee, Gwo-Bin [1 ,2 ,5 ]
机构
[1] Natl Cheng Kung Univ, Dept Engn Sci, Tainan 701, Taiwan
[2] Natl Cheng Kung Univ, Inst Nanotechnol & Microsyst Engn, Tainan 701, Taiwan
[3] Natl Cheng Kung Univ, Inst Basic Med Sci, Tainan 701, Taiwan
[4] Natl Cheng Kung Univ, Dept Parasitol, Tainan 701, Taiwan
[5] Ind Technol Res Inst, Med Elect & Device Technol Ctr, Hsinchu 31040, Taiwan
关键词
RNA extraction; Reverse transcription; Magnetic bead; Microfluidics; Magnetic separator; mu-TAS; MEMS; POLYMERASE-CHAIN-REACTION; PHENOL-CHLOROFORM EXTRACTION; SINGLE-STEP METHOD; RNA EXTRACTION; ENTEROVIRUS RNA; ON-CHIP; DNA; CELLS; SEPARATION; TISSUE;
D O I
10.1007/s10544-008-9240-1
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
This paper presents a new integrated microfluidic chip that automatically performs ribonucleic acid (RNA) extraction and reverse transcription (RT) processes. The microfluidic system consists of a microfluidic control module and a magnetic bio-separator. The microfluidic control module can perform pumping and mixing of small amount of fluids and subsequent purification and concentration of RNA samples by incorporating with the magnetic bio-separator consisting of 2-dimension twisted microcoils. Notably, the magnetic bio-separators are developed either to generate the required magnetic field to perform the separation of magnetic beads or to work as a micro-heater to control the temperature field for the following RT process. Experimental results show that the total RNA can be successfully purified and extracted by using magnetic beads and the subsequent RT processing of the RNA can be performed automatically. Total RNA is successfully extracted and purified from T98 cells utilizing the microfluidic system, which is comparable with the conventional methods. The whole automatic procedure of RNA sample extraction only takes 35 min, which is much faster than the conventional method (more than 2 h). As a whole, the developed microfluidic system may provide a powerful platform for rapid RNA extraction and RT processes for further biomedical applications.
引用
收藏
页码:339 / 350
页数:12
相关论文
共 35 条
[1]   Use of magnetic beads versus guanidium thiocyanate-phenol-chloroform RNA extraction followed by polymerase chain reaction for the rapid, sensitive detection of enterovirus RNA [J].
Beaulieux, F ;
See, DM ;
LeparcGoffart, I ;
Aymard, M ;
Lina, B .
RESEARCH IN VIROLOGY, 1997, 148 (01) :11-15
[2]  
Bird IM, 2005, METH MOLEC MED, V108, P139
[3]   A micromachined DNA manipulation platform for the stretching and rotation of a single DNA molecule [J].
Chiou, CH ;
Lee, GB .
JOURNAL OF MICROMECHANICS AND MICROENGINEERING, 2005, 15 (01) :109-117
[4]   ISOLATION OF BIOLOGICALLY-ACTIVE RIBONUCLEIC-ACID FROM SOURCES ENRICHED IN RIBONUCLEASE [J].
CHIRGWIN, JM ;
PRZYBYLA, AE ;
MACDONALD, RJ ;
RUTTER, WJ .
BIOCHEMISTRY, 1979, 18 (24) :5294-5299
[5]   An integrated microfluidic biochemical detection system for protein analysis with magnetic bead-based sampling capabilities [J].
Choi, JW ;
Oh, KW ;
Thomas, JH ;
Heineman, WR ;
Halsall, HB ;
Nevin, JH ;
Helmicki, AJ ;
Henderson, HT ;
Ahn, CH .
LAB ON A CHIP, 2002, 2 (01) :27-30
[6]   A new magnetic bead-based, filterless bio-separator with planar electromagnet surfaces for integrated bio-detection systems [J].
Choi, JW ;
Ahn, CH ;
Bhansali, S ;
Henderson, HT .
SENSORS AND ACTUATORS B-CHEMICAL, 2000, 68 (1-3) :34-39
[7]  
CHOMCZYNSKI P, 1993, BIOTECHNIQUES, V15, P532
[8]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[9]   The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on [J].
Chomczynski, Piotr ;
Sacchi, Nicoletta .
NATURE PROTOCOLS, 2006, 1 (02) :581-585
[10]   Magnetic bead handling on-chip: new opportunities for analytical applications [J].
Gijs, MAM .
MICROFLUIDICS AND NANOFLUIDICS, 2004, 1 (01) :22-40