High-speed RNA microextraction technology using magnetic oligo-dT beads and lateral magnetophoresis

被引:39
作者
Lee, Hwanyong [1 ]
Jung, Jinhee [1 ]
Han, Song-I [1 ]
Han, Ki-Ho [1 ]
机构
[1] Inje Univ, Sch Nano Engn, Gimhae 621749, Gyongnam, South Korea
关键词
MESSENGER-RNA; SOLID-PHASE; PURIFICATION; BLOOD; SYSTEM; SEPARATION; DIAGNOSIS; SILICA; CELLS; DNA;
D O I
10.1039/c005145d
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This paper presents a high-speed RNA microextractor for the direct isolation of RNA from peripheral blood lysate using magnetic oligo-dT beads. The extraction is achieved through lateral magnetophoresis, generated by a ferromagnetic wire array inlaid on a glass substrate. This RNA microextractor separated more than 80% of magnetic beads with a flow rate up to 20 ml h(-1), and the overall extraction procedure was completed within 1 min. The absorbance ratio of RNA to protein (A(260)/A(280)) was > 1.7, indicating that the extraction technology yielded nearly pure RNA. The feasibility of this technique was evaluated further for its applicability to reverse transcription polymerase chain reaction (RT-PCR) procedures by performing cDNA synthesis and PCR. The analysis verified that the RNA microextractor is a practical method for easy, rapid, and high-precision RT-PCR using minimal reagent volumes without requiring highly trained personnel. In addition, it can be readily incorporated into genetic analysis procedures for realizing automated on-chip genetic platforms in a micro format.
引用
收藏
页码:2764 / 2770
页数:7
相关论文
共 31 条
[21]   Microfluidic single-cell mRNA isolation and analysis [J].
Marcus, Joshua S. ;
Anderson, W. French ;
Quake, Stephen R. .
ANALYTICAL CHEMISTRY, 2006, 78 (09) :3084-3089
[22]   Parallel picoliter RT-PCR assays using microfluidics [J].
Marcus, JS ;
Anderson, WF ;
Quake, SR .
ANALYTICAL CHEMISTRY, 2006, 78 (03) :956-958
[23]   On-chip free-flow magnetophoresis: Separation and detection of mixtures of magnetic particles in continuous flow [J].
Pamme, Nicole ;
Eijkel, Jan C. T. ;
Manz, Andreas .
JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS, 2006, 307 (02) :237-244
[24]  
*QIAGEN, 2006, QIAAMP RNA BLOOD MIN
[25]  
Sambrook J., 2001, MOL CLONING A LAB MA, V3rd
[26]   Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains [J].
Setzer, Mindy ;
Juusola, Jane ;
Ballantyne, Jack .
JOURNAL OF FORENSIC SCIENCES, 2008, 53 (02) :296-305
[27]  
Takano T, 1998, CANCER RES, V58, P4913
[28]  
Wolfe KA, 2002, ELECTROPHORESIS, V23, P727, DOI 10.1002/1522-2683(200203)23:5<727::AID-ELPS727>3.0.CO
[29]  
2-O
[30]   Microchip-based macroporous silica sol-gel monolith for efficient isolation of DNA from clinical samples [J].
Wu, Qirong ;
Bienvenue, Joan M. ;
Hassan, Benjamin J. ;
Kwok, Yien C. ;
Giordano, Braden C. ;
Norris, Pamela M. ;
Landers, James P. ;
Ferrance, Jerome P. .
ANALYTICAL CHEMISTRY, 2006, 78 (16) :5704-5710