organism: Mus musculus
description: Protocols: Splenic lymphocytes were isolated from 7-9-week-old female wildtype (WT) mice and Itch knocked-out (KO) mice (3 mice/group) using Ficoll (Ficoll-Paque Plus, GE Healthcre). B cells were sorted by B220 microbeads (Cat No. 130-049-501, Miltenyi Biotec, Germany). Samples were washed twice with PBS and 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. Mice were euthanized using CO2. Spleens were cut using scissors. B cells were sorted by B220 microbeads (Cat No. 130-049-501, Miltenyi Biotec, Germany). B cells were not cultured in vitro. 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. Wildtype (WT) mice and Itch knocked-out (KO) mice were developed by Cyagen Co., Ltd (Guangzhou, China). Splenic lymphocytes were isolated from 7-9-week-old female wildtype (WT) mice and Itch knocked-out (KO) mice (3 mice/group) using Ficoll (Ficoll-Paque Plus, GE Healthcre). B cells were sorted by B220 microbeads (Cat No. 130-049-501, Miltenyi Biotec, Germany). Samples were washed twice with PBS and 1 mL of TRIzol per 1 x 107 cell pellets was added and frozen in -80℃. Data were aligned using STAR with options --outFilterScoreMinOverLread 0.4 --outFilterMatchNminOverLread 0.4, against the C3H/HeJ genome as a reference (ftp://ftp.ensembl.org/pub/release-89/fasta/mus_musculus_c3hhej/dna/Mus_musculus_c3hhej.C3H_HeJ_v1.dna_sm.toplevel.fa.gz). Total RNA was extracted from 107 PBMCs using TRIzol reagent (Life Technologies).Total RNA from cells was purified using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands). All RNA content in the samples was measured with a NanoDrop®ND-1000 spectrophotometer, and the RNA quality was assessed using RNA 6000 NanoChips with the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA, USA). Libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-03-01T04:03:07Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-08-23T14:41:39Z
attributes.tag: External Id
attributes.value: SAMEA5885222
attributes.tag: INSDC center name
attributes.value: Beijing Institute of Basic Medical Sciences
attributes.tag: INSDC first public
attributes.value: 2020-03-01T04:03:07Z
attributes.tag: INSDC last update
attributes.value: 2019-08-23T14:41:39Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8273:Sample 2
attributes.tag: age
attributes.value: 7 to 9
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: Itch KO
attributes.tag: individual
attributes.value: pool of 3 mice
attributes.tag: organism part
attributes.value: spleen
attributes.tag: sample name
attributes.value: E-MTAB-8273:Sample 2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: C57BL/6