run in TBE £0.5 at room temperature for 2 h at 150 V. The following two (Q/q) 27-bp unmethylated oligonucleotides were used: 5 0 -GATCCTTCGCCTAGGCTC(A/G)CAGCG CGGGAGCGA-3 0 . A methylated q probe (q*) was generated by incorporating a methylated cytosine at the mutated CpG site during oligonucleotide synthesis. Transient transfection assayThe constructs contained 578 bp from IGF2 intron 3 (nucleotides 2868-3446), followed by the IGF2 P3 promoter (nucleotides 2222 to þ45 relative to the start of transcription) 12 and a luciferase reporter. C2C12 myoblast cells were grown to approximately 80% confluence. Cells were transiently co-transfected with the firefly luciferase reporter construct (4 mg) and a Renilla luciferase control vector (phRG-TK, Promega; 80 ng) using 10 mg Lipofectamine 2000 (Invitrogen). Cells were incubated for 25 h before lysis in 100 ml Triton lysis solution. Luciferase activities were measured using the Dual-Luciferase Reporter Assay System (Promega). The results are based on four triplicate experiments using two independent plasmid preparations for each construct. Statistical analysis was done with an analysis of variance.

JAK2 exon 12 mutations define a distinctive myeloproliferative syndrome that affects patients who currently receive a diagnosis of polycythemia vera or idiopathic erythrocytosis.

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