Whole transcriptome RNA sequencing on bone marrow and peripheral blood samples from patients with acute myeloid leukemia at diagnosis or relapse.

One hundred cryopreserved bone marrow and peripheral blood samples from patients with acute myeloid leukemia (AML) with 10-90% blasts were selected from the biobank of the Department of Hematology of Leiden University Medical Center (LUMC). The AML cases cover all subtypes, and specifically include known subtype-defining balanced chromosomal translocations according to the WHO classification. The samples were obtained from 96 patients and include three pairs of de novo and relapsed AML and one pair of de novo and presumed therapy-related AML (tAML). Total RNA was isolated from mononuclear cells without prior enrichment for leukemic blasts. The quality and integrity of total RNA was checked and RNA libraries were prepared using the TruSeq RNA library preparation kit v2 (Illumina, San Diego, CA) in an ISO/IEC 17025-accredited protocol. This workflow started with enrichment of messenger RNA by oligo dT magnetic beads. After fragmentation, cDNA synthesis was performed, followed by adaptor ligation and PCR amplification. Paired-end sequencing with a read length of 126 bp was performed on an Illumina HiSeq 2500 v4 sequencer to at least 12.5 Gbp per sample. Image analysis, base calling, and quality check was performed with Illumina data analysis pipeline RTA v1.18.64 and Bcl2fastq v1.8.4. RNAseq reads are provided in compressed Sanger FASTQ format.

从莱顿大学医学院（LUMC）血液科生物库中选取了一百份急性髓系白血病（AML）患者的冷冻保存骨髓和外周血样本，其中 blast 细胞占 10-90%。这些 AML 病例涵盖了所有亚型，并特别包括根据世界卫生组织（WHO）分类已知的亚型定义性平衡染色体易位。这些样本来源于 96 名患者，包括三对原发性和复发性 AML 以及一对原发性和疑似治疗相关 AML（tAML）。从未经过白血病 blast 细胞富集的单核细胞中提取了总 RNA。通过检查总 RNA 的质量和完整性，并使用 Illumina 公司位于美国圣地亚哥的 TruSeq RNA 库制备试剂盒 v2（ISO/IEC 17025 认证协议）制备 RNA 库。该工作流程始于通过 oligo dT 磁性 beads 对信使 RNA 进行富集。经过断裂后，进行 cDNA 合成，随后进行接头连接和 PCR 扩增。在 Illumina HiSeq 2500 v4 测序仪上对配对末端测序进行了操作，读长为 126 碱基对，每个样本至少达到 12.5 Gbp。图像分析、基线读取和质量检查使用 Illumina 数据分析管道 RTA v1.18.64 和 Bcl2fastq v1.8.4 进行。RNA 测序读段以压缩的 Sanger FASTQ 格式提供。