Figure 1 | Blood Cancer Journal

Figure 1

From: Crizotinib treatment for refractory pediatric acute myeloid leukemia with RAN-binding protein 2-anaplastic lymphoma kinase fusion gene

Figure 1

(a) Sanger sequencing of RANBP2-ALK fusion transcripts. The breakpoint lies between RANBP2 exon 16 and ALK exon 20. (b) RT-PCR for RANBP2-ALK fusion transcripts in bone marrow aspirate samples. Total cellular RNA was extracted from leukemia blast cells with an RNeasy Mini Kit (QIAGEN, Tokyo, Japan). For RT-PCR analysis, 500 ng of total RNA was reverse-transcribed by PrimeScriptTM RT Master Mix according to the manufacturer’s instructions (TaKaRa Bio, Tokyo, Japan). PCR reactions contained cDNA template, TaKaRa Ex Taq (TaKaRa Bio), 10 × Ex Taq buffer (TaKaRa Bio), dNTPs (TaKaRa Bio), forward primer (5′-CATTCTACACCGTCTCCTACCAG-3′) and reverse primer (5′-CGAGGTGCGGAGCTTGCTCAGC-3′) in a 50 μl reaction.8 The cycling conditions were as follows: one cycle of 94 °C for 5 min; 30 cycles of 94 °C for 30 s, 60 °C for 30 s and 72 °C for 30 s; and one cycle of 72 °C for 7 min. Sequencing of the PCR product was performed by FASMAC Co., Ltd. (Kanagawa, Japan). CR, complete remission; HCT, hepatopoietic cell transplantation; RT-PCR, reverse transcription-PCR.

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