Comparison of the MagNA pure LC automated system and the RiboPure-blood RNA manual method for RNA extraction from multiple myeloma bone marrow samples conserved in an RNA stabilizer

被引:7
作者
Garcia-Effron, G. [1 ]
Gamarra, S. [1 ]
Crooke, A. [1 ]
Martinez-Sanchez, P. [1 ]
Lahuerta, J. [1 ]
Martinez-Lopez, J. [1 ]
机构
[1] Univ Hosp, Serv Hematol, Mol Biol Lab, Madrid, Spain
关键词
bone marrow; MagNA pure; multiple myeloma; RiboPure; RNA extraction;
D O I
10.1111/j.1751-553X.2006.00830.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
A total of 62 frozen bone marrow specimens conserved in RNA later (R) (Ambion) were processed using two different extraction methods, the MagNA Pure LC system (MAG; Roche) and the manual RiboPure-Blood RNA method (RIBO; Ambion); Beta glucoronidase RNA (GUS) was amplified by LightCycler PCR to evaluate the quality of both extraction procedures. Less than 1000 GUS copies/ml was detected in 26 of 62 specimens (41.94%) processed by MAG and in five of 62 specimens (8.06%) processed by RIBO. Moreover, RNA recovery from the 62 specimens by MAG is, on average, 2.91 cycle threshold-fold higher than RIBO (P = 0.0008). Furthermore, we compared the extraction times and reagent costs of both methods. In conclusion, RNA extraction using MAG is faster to process 32 samples and less expensive than RIBO but it is not sensitive enough to be employed for research purpose in our laboratory.
引用
收藏
页码:139 / 144
页数:6
相关论文
共 19 条
[1]   Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) - a Europe against cancer program [J].
Beillard, E ;
Pallisgaard, N ;
van der Velden, VHJ ;
Bi, W ;
Dee, R ;
van der Schoot, E ;
Delabesse, E ;
Macintyre, E ;
Gottardi, E ;
Saglio, G ;
Watzinger, F ;
Lion, T ;
van Dongen, JJM ;
Hokland, P ;
Gabert, J .
LEUKEMIA, 2003, 17 (12) :2474-2486
[2]  
BERILLONLAPOPIN L, 2001, BIOCHEMICA, V2, P9
[3]   Use of the MagNA pure LC automated nucleic acid extraction system followed by real-time reverse transcription-PCR for ultrasensitive quantitation of hepatitis C virus RNA [J].
Cook, L ;
Ng, KW ;
Bagabag, A ;
Corey, L ;
Jerome, KR .
JOURNAL OF CLINICAL MICROBIOLOGY, 2004, 42 (09) :4130-4136
[4]   Detection of herpes simplex virus DNA in genital and dermal specimens by LightCycler PCR after extraction using the IsoQuick, MagNA pure, and BioRobot 9604 methods [J].
Espy, MJ ;
Rys, PN ;
Wold, AD ;
Uhl, JR ;
Sloan, LM ;
Jenkins, GD ;
Ilstrup, DM ;
Cockerill, FR ;
Patel, R ;
Rosenblatt, JE ;
Smith, TF .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (06) :2233-2236
[5]   Urinary concentrating defect in mice with selective deletion of phloretin-sensitive urea transporters in the renal collecting duct [J].
Fenton, RA ;
Chou, CL ;
Stewart, GS ;
Smith, CP ;
Knepper, MA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (19) :7469-7474
[6]  
Grotzer MA, 2000, MED PEDIATR ONCOL, V34, P438, DOI 10.1002/(SICI)1096-911X(200006)34:6<438::AID-MPO12>3.0.CO
[7]  
2-Q
[8]   Entirely automated quantification of human immunodeficiency virus type 1 (HIV-1) RNA in plasma by using the ultrasensitive COBAS AMPLICOR HIV-1 monitor test and RNA purification on the MagNA pure LC instrument [J].
Hölzl, G ;
Stöcher, M ;
Leb, V ;
Stekel, H ;
Berg, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 2003, 41 (03) :1248-1251
[9]   Glycosylphosphatidylinositol-anchored ceruloplasmin is required for iron efflux from cells in the central nervous system [J].
Jeong, SY ;
David, S .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (29) :27144-27148
[10]  
Kessler HH, 2001, CLIN CHEM, V47, P1124