Molecular Monitoring of Chronic Myelogenous Leukemia

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Molecular Monitoring of Chronic Myelogenous Leukemia Y. Lynn Wang, Joong Won Lee, Ethel Cesarman, David K. Jin, Balazs Csernus The Journal of Molecular Diagnostics Volume 8, Issue 2, Pages 231-239 (May 2006) DOI: 10.2353/jmoldx.2006.040404 Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Schematic diagram of the two different sets of primers/probes for ABL quantification (left, ABL1; right, ABL2). Lightly shaded boxes represent ABL cDNA and darkly shaded boxes represent BCR cDNA with exons indicated. Numbers below cDNAs indicate nucleotide positions at exon boundaries. Arrows represent PCR primers and their relative positions to ABL and BCR-ABL cDNAs. Black bars represent the TaqMan probes and their positions. Sequences of primers and probes and their locations are shown under each diagram. The Journal of Molecular Diagnostics 2006 8, 231-239DOI: (10.2353/jmoldx.2006.040404) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Threshold cycles of the BCR-ABL and control genes in 12 pretreatment patient samples. Triplicate PCR reactions were performed for each patient sample. The mean of Ct values of 12 samples are represented by horizontal bars. The Journal of Molecular Diagnostics 2006 8, 231-239DOI: (10.2353/jmoldx.2006.040404) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Threshold cycle difference of six internal control genes in 10 pairs of pure patient samples and mixed samples. cDNA from 10 patient specimens (Table 1, patients 10 to 19) were mixed with cDNA from normal peripheral blood mononuclear cells at 1:16 ratio. Levels of six different control genes as indicated in the graphs were determined in the 10 pairs of pure and mixed samples. Mean Ct values of triplicate real-time PCR reactions were plotted. Lines connect pure and mixed patient samples to show pairwised relationship. The Journal of Molecular Diagnostics 2006 8, 231-239DOI: (10.2353/jmoldx.2006.040404) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 The degradation kinetics of four internal control transcripts in comparison with BCR-ABL. RNA was extracted from mononuclear cells from the bone marrow of two CML patients (Table 1, patients 20 and 21), placed at the bench for various periods of time. Real-time RT-PCR reactions were performed simultaneously for BCR-ABL and four different control genes as indicated in the legend boxes. Data at each time point after day 0 are expressed as a percentage of the initial time point calculated by the ΔCt method (for details, see Materials and Methods). Means ± SD are plotted. Note different y-axis scale is used in a and e. The Journal of Molecular Diagnostics 2006 8, 231-239DOI: (10.2353/jmoldx.2006.040404) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 The degradation kinetics of four internal control transcripts in comparison with BCR-ABL. RNA was extracted from mononuclear cells from the bone marrow of two CML patients (Table 1, patients 20 and 21), placed at the bench for various periods of time. Real-time RT-PCR reactions were performed simultaneously for BCR-ABL and four different control genes as indicated in the legend boxes. Data at each time point after day 0 are expressed as a percentage of the initial time point calculated by the ΔCt method (for details, see Materials and Methods). Means ± SD are plotted. Note different y-axis scale is used in a and e. The Journal of Molecular Diagnostics 2006 8, 231-239DOI: (10.2353/jmoldx.2006.040404) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions