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Amplification Refractory Mutation System, a Highly Sensitive and Simple Polymerase Chain Reaction Assay, for the Detection of JAK2 V617F Mutation in Chronic.

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Presentation on theme: "Amplification Refractory Mutation System, a Highly Sensitive and Simple Polymerase Chain Reaction Assay, for the Detection of JAK2 V617F Mutation in Chronic."— Presentation transcript:

1 Amplification Refractory Mutation System, a Highly Sensitive and Simple Polymerase Chain Reaction Assay, for the Detection of JAK2 V617F Mutation in Chronic Myeloproliferative Disorders  Qiaofang Chen, Pin Lu, Amy V. Jones, Nicholas C.P. Cross, Richard T. Silver, Y. Lynn Wang  The Journal of Molecular Diagnostics  Volume 9, Issue 2, Pages (April 2007) DOI: /jmoldx Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 Schematic diagram of the ARMS-PCR assay. Two forward and two reverse primers are used in different combinations to generate three potential PCR products. Primers FO and RO flank the exon 14 of the JAK2 gene, resulting in a band of 463 bp to control for DNA quality and quantity. Primers Fwt and RO amplify a wild-type allele, generating a band of 229 bp, and primers FO and Rmt generate a band of 279 bp from the mutant allele. Primer sequences are shown below the diagram. The intended mismatches in Fwt and Rmt are shown in lowercase, and genotype-specific nucleotides are underlined. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 The modified ARMS-PCR has a higher sensitivity than the original method. DNA samples from two patients were diluted with DNA from peripheral blood mononuclear cell at ratios of 1:10, 1:100, and 1:1000. The positions of the DNA control, mutant, and normal alleles are indicated. MW, 100-bp molecular weight markers. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

4 Figure 3 Analytic sensitivity of the ARMS-PCR assay. Genomic DNA from HEL cells was diluted with DNA from human peripheral blood mononuclear cell at indicated percentages. The positions of the DNA control, mutant, and normal alleles are indicated. MW, 100-bp molecular weight markers. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

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