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JMD 2006, Vol. 8, No. 3
Copyright © 2006 American Society for Investigative Pathology & Association for Molecular Pathology

Detection of the JAK2 V617F Mutation by LightCycler PCR and Probe Dissociation Analysis

Marla Lay*, Rajan Mariappan{dagger}, Jason Gotlib{ddagger}, Lisa Dietz*, Siby Sebastian§, Iris Schrijver*{dagger} and James L. Zehnder*{dagger}

From the Molecular Pathology Laboratory, * and Departments of Pathology {dagger} and Medicine (Hematology), {ddagger} Stanford University School of Medicine, Stanford Hospital and Clinics and Lucile Packard Children’s, Hospital, Stanford, California; and the Department of Pathology, § Duke University Medical Center, Durham, North Carolina

A point mutation in the JAK2 gene, a member of the tyrosine kinase family, was recently identified and shown to be associated with several myeloproliferative disorders. Several studies identified the same JAK2 point mutation (1849G>T), resulting in the substitution of a valine to phenylalanine at codon 617 (V617F). We developed a simple and sensitive method to detect this mutation via polymerase chain reaction and probe dissociation analysis using the LightCycler platform, and we compared this method to existing restriction fragment-length polymorphism, direct sequencing, and amplification refractory mutation system methods. We found that the LightCycler method offered advantages of speed, reliability, and more straightforward interpretation over the restriction fragment-length polymorphism and sequencing approaches.




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E. Hammond, K. Shaw, B. Carnley, S. P'ng, I. James, and R. Herrmann
Quantitative Determination of JAK2 V617F by TaqMan: An Absolute Measure of Averaged Copies per Cell That May Be Associated with the Different Types of Myeloproliferative Disorders
J. Mol. Diagn., April 1, 2007; 9(2): 242 - 248.
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