Validation of lab-on-chip capillary electrophoresis systems for total RNA quality and quantity control

被引:12
作者
Pfaffl, M. W. [1 ]
Fleige, S. [1 ]
Riedmaier, I. [1 ]
机构
[1] Tech Univ Munich, Ctr Life & Food Sci ZIEL, Physiol Weihenstephan, D-8050 Freising Weihenstephan, Germany
关键词
RNA integrity; RNA quality; RNA quantity; transcriptomics; lab-on-chip; capillary electrophoresis;
D O I
10.1080/13102818.2008.10817562
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Purity and good RNA quality are important elements for the overall success of RNA based analysis methods like microarrays and real tine qRT-PCR. There are two commercially available automated systems - the Experion (Bio-Rad Laboratories) and the 2100 Bioanalyzer (Agilent Technologies) - that provide both RNA sample quality and quantity analysis. In this study different aspects like the reproducibility, and sensitivity of both systems were analyzed by determining the total RNA quality and quantity\ extracted from various bovine tissues. Regarding quantitation, the Experion is more sensitive than the: 2100 Bioanalyzer: Both systems overstate the concentration by 19-29% compared to the photometric values. For RNA quality determination, both systems show highly comparable reproducibility. With the RNA integrity number (RIN) the 2100 Bioanalyzer offers an additional opportunity to quantify the RNA quality.
引用
收藏
页码:829 / 834
页数:6
相关论文
共 6 条
[1]   Chipping away at the chip bias: RNA degradation in microarray analysis [J].
Auer, H ;
Lyianarachchi, S ;
Newsom, D ;
Klisovic, MI ;
Marcucci, U ;
Kornacker, K .
NATURE GENETICS, 2003, 35 (04) :292-293
[2]   High quality RNA isolation from tumours with low cellularity and high extracellular matrix component for cDNA microarrays:: application to chondrosarcoma [J].
Baelde, HJ ;
Cleton-Jansen, AM ;
van Beerendonk, H ;
Namba, M ;
Bovée, JVMG ;
Hogendoorn, PCW .
JOURNAL OF CLINICAL PATHOLOGY, 2001, 54 (10) :778-782
[3]   Impact of RNA degradation on gene expression profiles:: Assessment of different methods to reliably determine RNA quality [J].
Copois, Virginie ;
Bibeau, Frederic ;
Bascoul-Mollevi, Caroline ;
Salvetat, Nicolas ;
Chalbos, Patrick ;
Bareil, Corinne ;
Candeil, Laurent ;
Fraslon, Caroline ;
Conseiller, Emmanuel ;
Granci, Virginie ;
Maziere, Pierre ;
Kramar, Andrew ;
Ychou, Marc ;
Pau, Bernard ;
Martineau, Pierre ;
Molina, Franck ;
Del Rio, Maguy .
JOURNAL OF BIOTECHNOLOGY, 2007, 127 (04) :549-559
[4]  
Fleige Simone, 2006, Molecular Aspects of Medicine, V27, P126, DOI 10.1016/j.mam.2005.12.003
[5]   Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR [J].
Fleige, Simone ;
Walf, Vanessa ;
Huch, Silvia ;
Prgomet, Christian ;
Sehm, Julia ;
Pfaffl, Michael W. .
BIOTECHNOLOGY LETTERS, 2006, 28 (19) :1601-1613
[6]  
MUELLER O, 2004, 5989 AG TECHN