title: LCK_6#61
organism: Danio rerio
description: Protocols: The spleen from a heterozygote Tg(lck:EGFP) or wild-type fish was 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: 2016-05-26
attributes.tag: External Id
attributes.value: SAMEA4012019
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: 2016-12-01T17:01:24Z
attributes.tag: INSDC last update
attributes.value: 2016-05-26T17:59:12Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-4617:LCK_6#61
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: zebrafish
attributes.tag: fsc
attributes.value: 19706
attributes.tag: gfp
attributes.value: 320
attributes.tag: individual
attributes.value: 2
attributes.tag: pi
attributes.value: 2
attributes.tag: plate
attributes.value: 6
attributes.tag: sample name
attributes.value: E-MTAB-4617:LCK_6#61
attributes.tag: ssc
attributes.value: 9384
attributes.tag: tissue
attributes.value: Spleen
attributes.tag: well
attributes.value: E8