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

International Journal of Laboratory Hematology - Tập 29 Số 2 - Trang 139-144 - 2007
Guillermo García‐Effrón1, Soledad Gamarra1, Almudena Crooke1, Pilar Martínez‐Sánchez1,2, Juan José Lahuerta2, Joaquín Martínez‐López1,2
1Laboratorio de Biología Molecular, Servicio de Hematología, Hospital Universitario, Madrid, Spain
2Servicio de Hematología, Hospital Universitario, Madrid, Spain

Tóm tắt

Summary

A total of 62 frozen bone marrow specimens conserved in RNA later® (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.

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Tài liệu tham khảo

10.1038/sj.leu.2403136

Bérillon‐Lapopin L., 2001, Evaluation of the MagNA Pure LC mRNA Isolation kit II (Tissue), Biochemica, 2, 9

10.1128/JCM.42.9.4130-4136.2004

10.1128/JCM.39.6.2233-2236.2001

10.1073/pnas.0401704101

10.1002/(SICI)1096-911X(200006)34:6<438::AID-MPO12>3.0.CO;2-Q

10.1128/JCM.41.3.1248-1251.2003

10.1074/jbc.M301988200

Kessler H.H., 2001, Fully automated nucleic acid extraction: MagNA Pure LC, Clinical Chemistry, 47, 1124, 10.1093/clinchem/47.6.1124

10.1128/JCM.42.2.585-590.2004

10.1128/JCM.40.6.2240-2243.2002

10.1373/49.6.955

10.1016/S0022-1759(03)00237-0

10.1128/JCM.41.8.3840-3845.2003

10.1186/1471-2164-5-88

10.1016/j.yexmp.2004.05.005

10.1128/JCM.43.9.4616-4622.2005

10.1016/j.jviromet.2005.05.002