title: CTL_RPF_1
organism: Mus musculus
description: Protocols: Meis1/Hoxa9-transduced foetal liver c-Kit+ cells were collected after three rounds of serial re-plating in methylcellulose. Cells were lysed with CaCl2 4 mM, MgCl 10 mM, K-HEPES pH 7.2 25 mM, KOAc 200 mM and NP-40 1%. Total RNA was extracted using QIAzol Lysis Reagent (QIAGEN). The lysate was cleared from cell debris, diluted 1:1 in water, and digested with MNase 10 μg/ml for 30 minutes at 37°C. Digested RNA was extracted with QIAzol and later treated with PNK (NEB) for 30 minutes at 37°C. To isolate ribosome-protected mRNA fragments (RPFs), the PNK-treated RNA was resolved on a 15% Novex TBE-Urea Gel (EC6885BOX), and RPFs 25 to 40 nucleotides long were excised and purified. Libraries were then prepared using the NEBNext® Multiplex Small RNA Library Prep Set for Illumina following manufacturer's instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-04-05T04:04:05Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-03-18T11:06:12Z
attributes.tag: External Id
attributes.value: SAMEA5425603
attributes.tag: INSDC center name
attributes.value: MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh
attributes.tag: INSDC first public
attributes.value: 2019-04-05T04:04:05Z
attributes.tag: INSDC last update
attributes.value: 2019-03-18T11:06:12Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7785:CTL_RPF_1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: pre-leukemic stem cell
attributes.tag: cell type
attributes.value: hematopoietic cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: embryonic day 14.5
attributes.tag: disease
attributes.value: acute myeloid leukemia
attributes.tag: fraction
attributes.value: ribosome protected fragments
attributes.tag: genotype
attributes.value: Ythdf2 fl/fl
attributes.tag: organism part
attributes.value: hematopoietic system
attributes.tag: sample name
attributes.value: E-MTAB-7785:CTL_RPF_1
attributes.tag: scientific_name
attributes.value: Mus musculus