|Values are valid only on day of printing.|
Total RNA is extracted from blood or bone marrow using the QIAmp kit (Qiagen). cDNA is made using the Superscript III kit (Invitrogen). Quantitative, real-time PCR is performed using the LightCycler instrument (Roche) and the data analyzed using the supplied software for relative quantification with calibrator normalization and efficiency correction.(Instruction manual: Roche Applied Science LightCycler 2.0 Instrument. Version 4/1.2. 2003. Roche Diagnostics)
The normalized ratio is a relative quantification calculation as follows:
*Where GUSB mRNA is used to normalize variations in RNA quality and calibrator mRNA from a promyelocytic leukemia/retinoic acid receptor alpha (PML/RARA)-positive cell line is used to normalize variations in run conditions.
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