PML/RARA Quantitative, PCR
Method Description Describes how the test is performed and provides a method-specific reference
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. (Roche Applied Science LightCycler 2.0 Instrument Operation Manual. 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 PML/RARA-positive cell line is used to normalize variations in run conditions.
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday