Illumina HiSeq 2500 sequencing; MicroRNA-sequencing of Brazilian pediatric acute lymphoblastic leukemia
MicroRNA-sequencing of Brazilian pediatric acute lymphoblastic leukemia
The leukemic samples were collected from the bone barrow aspirates of patients referred to the Hospital with suspect of leukemia. Bone marrow aspirates were used to separation of the mononuclear cells for morphological classication of blasts (FAB criteria) and immunophenotyping analysis (flow citometry) for precise leukemia subtype definition. The same mononuclear fraction (an aliquot) was used to the microRNA sequencing experiments and analysis.  We performed total RNA extraction using the TRIzol reagent (Invitrogen, Carlsbad, California), following the manufacturer's instructions. miRNA library construction was performed starting from 1 Î¼g of each RNA sample using the TruSeq Small RNA Library Prep kit (Illumina, San Diego, California), following manufacturer's instructions.