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Detection of the JAK2 V617F mutation by LightCycler PCR and probe dissociation analysis.


ABSTRACT: 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 (1,849G>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.

SUBMITTER: Lay M 

PROVIDER: S-EPMC1867612 | biostudies-literature | 2006 Jul

REPOSITORIES: biostudies-literature

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Detection of the JAK2 V617F mutation by LightCycler PCR and probe dissociation analysis.

Lay Marla M   Mariappan Rajan R   Gotlib Jason J   Dietz Lisa L   Sebastian Siby S   Schrijver Iris I   Zehnder James L JL  

The Journal of molecular diagnostics : JMD 20060701 3


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 (1,849G>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  ...[more]

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