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PCR experiments were performed starting from 100 ng of randomly primed reverse-transcribed RNA for 35 cycles. The primers used for PCR were B8, B12 (25), P3 (tggaattcTGCAGTGGAGGAGATAAA), and P6 (CCTTGCCAAGTTGCTGTAGA) for JAK2. The specificity of the amplifications was verified by nucleotide sequence analysis.
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4 cells per well). The number of wells showing proliferating cells in either the absence or presence of IL-3 was scored after 1 week in culture.
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The cDNA from patient 2 (an atypical chronic myelogenous leukemia case in transformation) was amplified with the TEL primer ATGCACCCTCTGATCCTGAACC and the JAK2 primer TGGTGAGGTTGGTACATCAG. The cDNA from patient 3 (a precursor B cell ALL case) was amplified with the TEL primer TTCCACCCTGGAAACTCTATA and the JAK2 primer AAGGTTTGCTAATTCTGCCCACTTTGGTGC. PCR products were sequenced on an ALF sequencer.
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note
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We thank M. Le Coniat for FISH experiments, H. Beug for the murine Jak2 cDNA, P. Mayeux for helpful hints, P. Peeters and P. Marynen for sharing unpublished data, and S. Gisselborecht and G. Calothy for critical reading of the manuscript. Supported by the Association pour la Recherche sur le Cancer (V.L. and A.B.), the Ligue Nationale Contre le Cancer (C.T.Q.), and funds from INSERM, CNRS, Institut Curie, Association pour la Recherche sur le Cancer, Ligue Nationale Contre le Cancer, Ligue Départementale Contre le Cancer (Comité de Paris), and the Biomed Programme of the European Community (contract BMH4-CT96-1355).
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