organism: Mus musculus
description: Protocols: Mice kidneys were snap-frozen after nephrectomy. RNeasy mini Kit (Qiagen, Hilden, Germany) was used to isolate RNA from snap-frozen kidneys as per manufacturer's instructions. Ribosomal RNA was removed by using biotinylated target-specific oligos combined with Ribo-Zero rRNA removal beads from 1µg totalRNA input. The Ribo-Zero Human/Mouse/Rat kit depletes samples of cytoplasmic rRNA. Following purification, the RNA is fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments then have the addition of a single'A' base and subsequent ligation of the adapter. The products are purified and enriched with PCR (20ul template, 15cycles) to create the final cDNA library. After library validation and quantification, equimolar amounts of library were pooled. Pools of 5-6 libraries each were quantified by using the Peqlab KAPA Library Quantification Kit and the Applied Biosystems 7900HT Sequence Detection System and sequenced on Hiseq 2000.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-01-03T16:00:55Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-05-21T13:11:00Z
attributes.tag: External Id
attributes.value: SAMEA5624881
attributes.tag: INSDC center name
attributes.value: Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne
attributes.tag: INSDC first public
attributes.value: 2020-01-03T16:00:55Z
attributes.tag: INSDC last update
attributes.value: 2019-05-21T13:11:00Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7982:HP12
attributes.tag: age
attributes.value: 8 to 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: diet
attributes.value: ad libitum
attributes.tag: growth condition
attributes.value: hypoxia
attributes.tag: injury
attributes.value: no ischemia-reperfusion injury
attributes.tag: organism part
attributes.value: kidney
attributes.tag: sample name
attributes.value: E-MTAB-7982:HP12
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL6/J