Garcia-Effron G, Gamarra S, Crooke A, Martínez-Sánchez P, Lahuerta J, Martínez-López J
Laboratorio de Biología Molecular, Servicio de Hematología, Hospital Universitario, Madrid, Spain.
Int J Lab Hematol. 2007 Apr;29(2):139-44. doi: 10.1111/j.1751-553X.2006.00830.x.
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.
总共62份保存在RNA later(安必昂公司)中的冷冻骨髓标本,采用两种不同的提取方法进行处理,即MagNA Pure LC系统(MAG;罗氏公司)和手动RiboPure - Blood RNA方法(RIBO;安必昂公司);通过LightCycler PCR扩增β - 葡萄糖醛酸酶RNA(GUS),以评估两种提取方法的质量。在MAG处理的62份标本中的26份(41.94%)以及RIBO处理的62份标本中的5份(8.06%)中,检测到GUS拷贝数低于1000拷贝/毫升。此外,MAG从62份标本中回收的RNA平均比RIBO高2.91个循环阈值倍数(P = 0.0008)。此外,我们比较了两种方法的提取时间和试剂成本。总之,使用MAG提取RNA处理32个样本更快,且比RIBO成本更低,但在我们实验室中其灵敏度不足以用于研究目的。