title: LCK_8#2
organism: Danio rerio
description: Protocols: Spleens from two heterozygote Tg(lck:EGFP) fish were dissected and passed through a 40μm cell strainer using the plunger of a 1-mL syringe and cells were collected in cold 1xPBS/5% FBS. A non-transgenic line was used to set up the gating and exclude autofluorescent cells. Propidium iodide (PI) staining was used to exclude dead cells. Individual cells were sorted, using a Becton Dickinson Influx sorter with 488- and 561 nm lasers (Schulte et al., 2015) and collected in a single well of a 96 well plate containing 2.3 uL of 0.2 % Triton X-100 supplemented with 1 U/uL SUPERase In RNAse inhibitor (Ambion). The size, granularity and level of fluorescence for each cell were simultaneously recorded. The Smart-seq2 protocol (Picelli et al., 2014) was used to amplify the whole transcriptome and prepare libraries. Twenty-five cycles of PCR amplification were performed.
attributes.tag: ENA first public
attributes.value: 2016-12-01
attributes.tag: ENA last update
attributes.value: 2016-09-12
attributes.tag: External Id
attributes.value: SAMEA4432091
attributes.tag: INSDC center alias
attributes.value: Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland Swiss Institute of Bioinformatics
attributes.tag: INSDC center name
attributes.value: Ludwig Center for Cancer Research, University of Lausanne, Lausanne, Switzerland Swiss Institute of Bioinformatics
attributes.tag: INSDC first public
attributes.value: 2016-12-01T17:01:25Z
attributes.tag: INSDC last update
attributes.value: 2016-09-12T14:14:53Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: PI fluorescence
attributes.value: 15
attributes.tag: Submitter Id
attributes.value: E-MTAB-46171473686057:LCK_8#2
attributes.tag: age
attributes.value: 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: zebrafish
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: forward scatter
attributes.value: 38983
attributes.tag: genotype
attributes.value: heterozygote Tg(lck:EGFP)
attributes.tag: gfp fluorescence
attributes.value: 287
attributes.tag: individual
attributes.value: 2
attributes.tag: organism part
attributes.value: spleen
attributes.tag: plate
attributes.value: 8
attributes.tag: sample name
attributes.value: E-MTAB-46171473686057:LCK_8#2
attributes.tag: side scatter
attributes.value: 51
attributes.tag: single cell well quality
attributes.value: good quality
attributes.tag: well
attributes.value: A2
attributes.tag: well information
attributes.value: single cell