mill Applications in Oncology
AML 1
ETO
F
3
4
3
4
5
:7
R
AML 1-ETO fusion trooscriIX
Fig. I. Schematic diagram of the AMLl-ETO transcript with AMLl exon 5 fused to ETO exon 2.
The breakpoint region and the position of the forward (F) and reverse (R) primers and the
hybridization probes are indicated
established a sensitive method to accurately quantify minimal residual leukemia.
Quantification of AMLl-ETO transcript levels may lead to the detection of a critical MRD level or via disease kinetics (increase and decrease of transcript levels) reveal patients with a high risk of hematological relapse before clinical
manifestation.
Materials
Equipment LightCycler instrument (Roche Diagnostics, Mannheim, Germany)
LightCycler software 3.0 and a preliminary-version of the relative quantification
software (kindly provided by Dr. R. Soong, Roche Diagnostics, Penzberg)
LightCycler capillaries and centrifuge (Roche Diagnostics)
EDTA stabilized human blood and bone marrow samples
Kasumi 1 cell line [8].
Reagents LightCycler Faststart DNA Master Hybridization Probes (Roche Diagnostics)
1st Strand cDNA Synthesis Kit for RT-PCR (AMV) (Boehringer, Mannheim,
Germany)
Qiagen RNeasy RNA-Isolation Kit (Qiagen, Hilden, Germany)
Hybridization Probes (TIB MOLBIOL, Berlin, Germany)
Primers (TIB MOLBIOL)
Ficoll Hypaque density gradient (Sigma, Munich, Germany)
AML 1
ETO
F
3
4
3
4
5
:7
R
AML 1-ETO fusion trooscriIX
Fig. I. Schematic diagram of the AMLl-ETO transcript with AMLl exon 5 fused to ETO exon 2.
The breakpoint region and the position of the forward (F) and reverse (R) primers and the
hybridization probes are indicated
established a sensitive method to accurately quantify minimal residual leukemia.
Quantification of AMLl-ETO transcript levels may lead to the detection of a critical MRD level or via disease kinetics (increase and decrease of transcript levels) reveal patients with a high risk of hematological relapse before clinical
manifestation.
Materials
Equipment LightCycler instrument (Roche Diagnostics, Mannheim, Germany)
LightCycler software 3.0 and a preliminary-version of the relative quantification
software (kindly provided by Dr. R. Soong, Roche Diagnostics, Penzberg)
LightCycler capillaries and centrifuge (Roche Diagnostics)
EDTA stabilized human blood and bone marrow samples
Kasumi 1 cell line [8].
Reagents LightCycler Faststart DNA Master Hybridization Probes (Roche Diagnostics)
1st Strand cDNA Synthesis Kit for RT-PCR (AMV) (Boehringer, Mannheim,
Germany)
Qiagen RNeasy RNA-Isolation Kit (Qiagen, Hilden, Germany)
Hybridization Probes (TIB MOLBIOL, Berlin, Germany)
Primers (TIB MOLBIOL)
Ficoll Hypaque density gradient (Sigma, Munich, Germany)